Possible role of the nipah virus v protein in the regulation of the interferon beta induction by interacting with ubx domain-containing protein1

ABSTRACT Nipah virus (NiV) is a highly pathogenic paramyxovirus that causes lethal encephalitis in humans. We previously reported that the V protein, one of the three accessory proteins

encoded by the P gene, is one of the key determinants of the pathogenesis of NiV in a hamster infection model. Satterfield B.A. _et al_. have also revealed that V protein is required for the

pathogenicity of henipavirus in a ferret infection model. However, the complete functions of NiV V have not been clarified. In this study, we identified UBX domain-containing protein 1

(UBXN1), a negative regulator of RIG-I-like receptor signaling, as a host protein that interacts with NiV V. NiV V interacted with the UBX domain of UBXN1 via its proximal zinc-finger motif

in the C-terminal domain. NiV V increased the level of UBXN1 protein by suppressing its proteolysis. Furthermore, NiV V suppressed RIG-I and MDA5-dependent interferon signaling by

stabilizing UBXN1 and increasing the interaction between MAVS and UBXN1 in addition to directly interrupting the activation of MDA5. Our results suggest a novel molecular mechanism by which

the induction of interferon is potentially suppressed by NiV V protein via UBXN1. SIMILAR CONTENT BEING VIEWED BY OTHERS NIPAH VIRUS W PROTEIN HARNESSES NUCLEAR 14-3-3 TO INHIBIT

NF-ΚB-INDUCED PROINFLAMMATORY RESPONSE Article Open access 16 November 2021 IKKΕ ISOFORM SWITCHING GOVERNS THE IMMUNE RESPONSE AGAINST EV71 INFECTION Article Open access 02 June 2021 THE E3

LIGASE ASB3 DOWNREGULATES ANTIVIRAL INNATE IMMUNITY BY TARGETING MAVS FOR UBIQUITIN-PROTEASOMAL DEGRADATION Article 12 September 2024 INTRODUCTION Nipah virus (NiV) is an emerging zoonotic

virus which belongs to the genus _Henipavirus_ in the family _Paramyxoviridae_, and was first identified as the pathogen that caused an outbreak of fatal encephalitis in humans in Malaysia

and Singapore in 19991,2. Since 2001, outbreaks of NiV have occurred sporadically almost every year in India, Bangladesh, and the Philippines3,4,5,6,7, and high mortality rates of around

40–70% in humans have been reported4,7. Serological evidence of henipavirus infections among humans and bats has also been reported in regions where no outbreaks have yet occurred8,9,10,11,

implying that other areas are threatened with future outbreaks of henipavirus infections. Therefore, it is important to understand the molecular mechanism underlying the severe pathogenesis

of henipavirus infections to allow the development of effective treatments. The genome of NiV is a negative-strand nonsegmented RNA containing six genes encoding structural proteins:

nucleocapsid (N), phosphoprotein (P), matrix protein (M), fusion protein (F), glycoprotein (G), and polymerase (L). The P gene encodes three additional accessory proteins: V, W, and C

proteins2,12. The RNA of the P gene is edited to generate the V and W genes, when additional guanines are inserted at the RNA editing site due to the stuttering of the RNA polymerase during

mRNA transcription12. Thus, the P, V, and W proteins share a common N-terminal domain, but each has a unique C-terminal domain. The C protein is encoded in an alternative reading frame of

the P gene. We have previously reported that the V and C proteins play key roles in the severe pathogenicity of NiV in a hamster infection model13. Satterfield B. A. _et al_. have also

reported that V protein is a determinant of the disease course of NiV infection, and C protein contributes to the respiratory disease in a ferret infection model14,15. Mathieu C. _et al_.

have revealed that C protein contributed to the virulence by regulating the early host proinflammatory response16. Recently, Marsh G.A. _et al_. isolated Ceder virus (CedPV) from Australian

bats as a novel member of henipaviruses. CedPV lacked the RNA editing site for the V protein expression, and was attenuated in ferrets and guinea pigs which are susceptible to NiV and Hendra

virus (HeV)17. It also indicates that V protein is a critical for the pathogenesis of henipaviruses. Previous studies have demonstrated that henipavirus V proteins suppress the host

antiviral response by targeting multiple host proteins. Henipavirus V proteins interact with MDA5 to inhibit the activation of the interferon β (_IFNB_) promoter18,19. NiV V binds to the

phosphatase PP1 to suppress the dephosphorylation of MDA5 and thereby its activation20. Henipavirus V proteins also interact with LGP2 to suppress the RIG-I-dependent induction of IFN21. NiV

V also blocks signaling through Toll-like receptors 7/9 (TLRs 7/9) and inhibitor of κB kinase ε to suppress IFN induction22,23. V proteins of NiV and HeV interacts with signal transducer

and activator of transcription 1 and 2 (STAT1 and STAT2) in IFN-responsive signaling pathway, through its N-terminal domain, which it shares with the P and W proteins, and prevents their

activation and nuclear accumulation24,25. During viral replication, NiV V negatively regulates minigenome replication26. These reports have revealed that V protein interacts with multiple

host proteins to contribute to the severe pathogenicity of henipavirus, and to elucidate the unknown molecular mechanism underlying the pathogenicity of henipavirus, the host molecule

interacting with henipavirus V proteins should be further investigated. In this study, we looked for unknown host proteins that interact with NiV V and identified UBX domain-containing

protein 1 (UBXN1). We then analyzed the effects of the interaction between NiV V and UBXN1. RESULTS IDENTIFICATION OF A PROTEIN THAT INTERACTS WITH NIV V PROTEIN To identify host proteins

that specifically interact with the unique C-terminal domain of NiV V, which is not shared with the P or W protein, myc-tagged wild-type NiV V and a mutant lacking the C-terminal domain

(ΔCT) were expressed in HEK293T cells and immunoprecipitated with an anti-myc antibody. Two protein bands coimmunoprecipitated with wild-type NiV V, but not with ΔCT were detected

(Supplementary Fig. 1A). One of the proteins with a mass of approximately 42.56 kDa was identified as UBX domain-containing protein 1 (UBXN1) by a mass-spectrometric analysis (Fig. 1A). To

confirm the result of the mass-spectrometric analysis, the same immunoprecipitation assay was followed by western blotting with an anti-myc antibody and an anti-UBXN1 antibody. UBXN1 was

detected as the protein binding to wild-type NiV V, but not to ΔCT (Fig. 1B). The interaction was also verified by the coimmunoprecipitation of NiV V with the endogenous UBXN1 (Fig. 1C). In

HeLa cells, UBXN1 was also coimmunoprecipitated with NiV V, indicating that UBXN1 is a binding partner of NiV V in various cell types (Fig. 1D). To confirm the requirement of the C-terminal

zinc-finger domain for the interaction, myc-tagged wild-type NiV V or ΔCT were coexpressed with HA-tagged UBXN1 in HEK293T cells and immunoprecipitated with either anti-myc antibody or

anti-HA antibody. The wild-type NiV V, but not ΔCT, was coimmunoprecipitated with UBXN1 (Fig. 1E). To confirm their intracellular interaction, an immunofluorescence assay was also performed

after NiV V or UBXN1 was expressed in HEK293T cells. As previously reported, NiV V was mainly localized in cytoplasm (Fig. 1F, upper lane)24. UBXN1 was also localized in cytoplasm, and

formed spot-like structures (Fig. 1F, middle lane), which is reportedly attributable to its association with the ubiquitin–proteasome system27,28. The coexpression of both proteins caused

the accumulation of UBXN1 in the cytoplasm, and its signal localized strongly with that of the V protein (Fig. 1F, lower lane). Furthermore, NiV V was co-localized with endogenous UBXN1 in

the cytoplasm (Fig. 1G). These results suggest that V protein interacts intracellularly with UBXN1. IDENTIFICATION OF THE BINDING DOMAINS OF NIV V AND UBXN1 The V protein contains two

cysteine-rich zinc-finger motifs in its C-terminal domain (Fig. 2A), which are conserved among the paramyxoviruses29. To determine the binding domain of NiV V, the C-terminal domain was

divided into three subdomains (Domain1–3) and vectors expressing various deletion mutants lacking each subdomain of NiV V (Del1–6), were generated (Fig. 2B). In an immunoprecipitation assay,

the binding affinity of NiV V for UBXN1 was detected with Del1 and Del5, but not with Del2, -3, -4, or -6 (Fig. 2C). The results indicate that Domain1 is critical for the interaction of the

two proteins. However, because Del2 did not coimmunoprecipitate with UBXN1, and the binding capacity of Del5 for UBXN1 was as strong as that of wild-type NiV V protein, whereas that of Del1

was much weaker, the intact structure of the proximal zinc-finger motif, including Domain1 and Domain3, must be important for the efficient interaction of the V protein with UBXN1. It has

been reported that UBXN1 consists of three characterized domains, a ubiquitin-associated (UBA) domain, a coiled-coil (CC) domain, and a ubiquitin regulatory X (UBX) domain27,28. To determine

the V-binding domain of UBXN1, expression vectors for GST-tagged deletion mutants of UBXN1 were generated: GST-CC, -UBA, -UBX, -ΔUBX, and -ΔCC, (Fig. 2D). In a GST pull-down assay, the V

protein was pulled down by two mutants of UBXN1, GST-UBX and -ΔCC, both of which contained the UBX domain, but not by the other mutants (Fig. 2E). This result suggests that NiV V interacts

with the UBX domain of UBXN1. NIV V INCREASES THE STABILITY OF UBXN1 We transfected the vector expressing HA-tagged UBXN1 together with that expressing NiV V or NiV P. The expression of

UBXN1 protein increased, depending on the amount of the NiV V expression vector transfected, but not for NiV P (Fig. 3A). The expression of enhanced green fluorescent protein (EGFP) was not

affected by the NiV V expression (Fig. 3B), indicating that NiV V specifically increased the expression of UBXN1. To examine the possibility that a high quantity of UBXN1 results in the

increase of its expression amount, we generated the HEK293 cell lines whose UBXN1 gene was knocked-out by a genome editing (Fig. 3C). As previously reported, the induction of IFNβ was

increased by the depletion of UBXN1 in both cell lines (Fig. S5A,B). We found that NiV V increased the expression amount of UBXN1 in 293TUBXN1− cells, indicating that the effects of NiV V on

the UBXN1 expression is not due to the artifacts derived from a high quantity of UBXN1 (Fig. 3D). Furthermore, this functional effect of NiV V was also observed in various cell types

including Huh-7 and HeLa cells (Fig. 3D). Since the mRNA levels of UBXN1 were not affected by NiV V (data not shown), we speculated that NiV V blocks the degradation of UBXN1. Therefore, a

cycloheximide (CHX) chase assay was performed to evaluate the rate of UBXN1 proteolysis when the _de novo_ protein synthesis was blocked. When NiV V was expressed together with UBXN1, the

degradation of UBXN1 was inhibited (Fig. 3E,F). This result suggests that NiV V increases the stability of UBXN1, causing its intracellular accumulation. STABILIZATION OF UBXN1 REQUIRES THE

INTERACTING DOMAINS OF NIV V AND UBXN1 To determine the domain of V required for the stabilization of UBXN1, the deletion mutants of NiV V described in Fig. 2B was expressed together with

UBXN1. The mutants containing Domain1 (Del1, -2 and -5) clearly increased the expression level of UBXN1, whereas the others did not (Fig. 4A). To determine the domain of UBXN1 required for

its stabilization by NiV V, we generated various vectors expressing HA-tagged deletion mutants of UBXN1, ΔUBX, ΔUBA, UBX, ΔCC, and CC (Fig. 4B). These mutants were expressed with or without

NiV V. The expression levels of the deletion mutants containing the UBX domain (ΔUBA, UBX, and ΔCC) increased when coexpressed with NiV V (Fig. 4C), but the others did not. These results

indicate that the stabilization of UBXN1 requires Domain1 of NiV V and the UBX domain of UBXN1, which are identical to the domains required for the interaction of the two proteins (Fig. 

2C,E). Therefore, we infer that the stabilization of UBXN1 is caused by its interaction with NiV V. AMINO ACIDS OF NIV V REQUIRED FOR ITS INTERACTION WITH UBXN1 Ramachandran and Horvath

reported that Domain1 of the V protein contains the amino acids required for MDA5 interference29. In this study, we found that Domain1 of the V protein also interacts with UBXN1 (Figs 2C and

5A), so we considered whether the same region in Domain1 functions in both MDA5 interference and the interaction with UBXN1. To evaluate the amino-acid requirements for these two functions,

we generated vectors expressing various myc-tagged alanine-substitution mutants of NiV V, RRE, ISI, CWD, GKR, AWV, and EEW (Fig. 5A). The mutants were expressed in HEK293T cells and an

immunoprecipitation assay was performed. Mutants RRE and EEW maintained the interaction with UBXN1, although it was weaker than that of intact NiV V. However, the interactions of mutants

ISI, CWD, GKR, and AWV were eliminated (Fig. 5B). To confirm the stabilization of UBXN1 by the myc-tagged alanine-substitution mutants of NiV V, these mutants were coexpressed with HA-tagged

UBXN1. RRE stabilized UBXN1 as effectively as did wild-type NiV V, but GKR and EEW stabilized it less effectively, and ISI, CWD, and AWV did not stabilize it at all (Fig. 5C). Therefore,

residues ISI, CWD, and AWV are important for UBXN1 stabilization. To determine the amino acids required for MDA5 interference, the alanine-substitution mutants of NiV V were coexpressed with

MDA5 in HEK293T cells, and the induction of IFN was monitored with an IFNβ reporter vector. MDA5 induced the activation of the _IFNB_ promoter, and wild-type NiV V interrupted MDA5 activity

(Fig. 5D). The alanine-substitution mutants CWD, GKR, and EEW reduced the activity of MDA5 as strongly as did wild-type NiV V, whereas the interference activities of RRE, ISI, and AWV were

weaker than that of wild-type NiV V (Fig. 5D). These results suggest that MDA5 and UBXN1 share the amino acid residues in ISI and AWV to interact with NiV V. On the other hand, CWD and GKR

contains the amino acid residues specifically critical for the interaction with UBXN1, and RRE contains those for the interaction with MDA5. Since the binding sites to UBXN1 and MDA5 in NiV

V were close, we evaluated the competitive binding of these proteins with immunoprecipitation assay. Although the coimmunoprecipitation of UBXN1 and MDA5 was slightly decreased in the

competitive binding, both proteins were coimmunoprecipitated with NiV V together (Fig. 5E). These results indicated that the binding sites of NiV V to MDA5 and UBXN1 were close but not

identical, and NiV V could interact with both proteins together. STABILIZED UBXN1 SUPPRESSES IFN INDUCTION UBXN1 is reportedly induced in a late step of viral infection, and binds to MAVS,

and this binding downregulates the induction of IFN by disrupting the MAVS–TRAF3/6 signaling complex in RIG-I-like receptor signaling30. NiV V is known to directly interfere with MDA5, thus

suppressing the induction of IFN18,21. To examine whether the stabilized UBXN1 increases suppression activity of NiV V on MDA5 signaling, IFN reporter assay was performed. The coexpression

of UBXN1 increased MDA5-suppression activity of NiV V, depending on the expression level (Fig. 6A). To examine whether RIG-I signaling was suppressed by the stabilized UBXN1, the

constitutively activated RIG-I (RIG-IΔ) was expressed with NiV V and UBXN1, and IFN reporter assay was performed. As previously reported19, NiV V did not suppress RIG-I signaling directly

(Fig. 6B). However, when coexpressed with UBXN1, NiV V suppressed RIG-I signaling depending on the expression level (Fig. 6B). This suppression effect of NiV V was also observed in 293UBXN1-

cells, indicating that it is not due to the artifacts derived from a high amount UBXN1 (Fig. 6C). The immunoprecipitation assay revealed that NiV V increased the interaction between UBXN1

and endogenous MAVS (Fig. 6D). These results suggest that NiV V suppresses the induction of IFN via RIG-I-like receptor signaling by stabilizing UBXN1 (Fig. 6E). DISCUSSION In this study, we

newly identified UBXN1 as a protein that interacts with NiV V, and demonstrated that they interact via Domain1 of NiV V and the UBX domain of UBXN1. NiV V increases the stability of UBXN1

by blocking its proteolysis, which requires the interacting domains of both proteins. With alanine-substitution mutants, we showed that Domain1 of NiV V contains the amino acids required

both for UBXN1 stabilization and MDA5 interference, but they are not identical. Our results showed that UBXN1 increased the suppression activity of NiV V on MDA5 activation, and conferred

the suppression activity on RIG-I activation to NiV V. These results potentially suggest a novel molecular mechanism for the suppression of IFN induction, in which NiV V stabilizes the

negative regulator of RIG-I-like receptor signaling, UBXN1. After recognizing viral RNA in the cytosol, RIG-I and MDA5 recruit MAVS via the CARD–CARD homotypic interaction31. The CARD domain

of MAVS rapidly forms prion-like aggregates, which interact with downstream signaling proteins, including tumor necrosis factor (TNF) receptor associated factors (TRAFs), resulting in the

activation of IKK and TBK132. Wang _et al_. reported that UBXN1 is induced in a late step of viral infection, and negatively regulates RIG-I-like receptor signaling, interacting with the

TRAF 3/6-binding site in MAVS through its N-terminal UBA domain as a dominant-negative binder, and thus disrupting MAVS–MAVS aggregation30. Our results indicate that NiV V interacts with the

C-terminal UBX domain of UBXN1 to stabilize it, suggesting that NiV V potentially enhances the negative regulation of RIG-I-like receptor signaling by stabilizing the induced UBXN1 during

viral infection. It has been reported that the V proteins of the paramyxoviruses do not interrupt MAVS directly22, but we have shown that NiV V could indirectly interrupts MAVS by

stabilizing its dominant-negative binding partner UBXN1. To the best of our knowledge, this is the first report of the interference of MAVS by the V proteins of the paramyxoviruses. It has

been reported that V protein targets multiple host proteins to suppress host’s IFN system18,20,21,22,23,24,25,33,34,35, and therefore we speculate that the ability of V protein to stabilize

UBXN1 potentially functions cooperatively with its other abilities such as the direct inhibition to MDA5, LGP2 and STAT proteins to suppress RIG-I- and MDA5-dependent IFN induction and

contribute to the pathogenesis. Our results showed that NiV V increased the amount of UBXN1 expressed by the plasmid transfection, but we could not recognize the increase of endogenous UBXN1

when NiV V was expressed. Our results also indicated that NiV V binds to UBX domain in UBXN1, and it has been reported that multiple host’s proteins such as BRCA1, Homer2 and p97 also bind

to UBX domain in UBXN127,28,36. Therefore, we speculate that the only small amount of endogenous UBXN1 in the cells interacted with NiV V because it had already bound to other host proteins,

which did not allow us to recognize the functional effects of NiV V on endogenous UBXN1. On the other hand, exogenously expressed UBXN1 might efficiently bind to NiV V before it binds to

other host’s proteins, which allow us to clearly show the functional effects of NiV V. It has been reported that the gene expression of UBXN1 is induced by the infection of RNA viruses30,

which suggests that NiV V potentially interacts with newly synthesized UBXN1 efficiently, and stabilizes it to suppress IFN induction. It is known that V proteins in paramyxoviruses shares

the zinc-finger structure in their C-terminal domains, and interact with common host proteins such as MDA5, LGP2 and STAT proteins18,19,37,38,39,40,41,42,43. Therefore, paramyxovirus V

proteins may also share the affinity to UBXN1. Since the Domain1 in the C-terminal domain of NiV V was identified as a binding site for UBXN1 (Figs 2C and 4A), we evaluated the homology of

amino acid sequence in the Domain1 among paramyxoviruses (Supplementary Table 1). The amino acid sequence of HeV V protein was almost identical to that of NiV V (identity: 89%), whereas

those of other genera including measles virus (MeV), mumps virus (MuV), Newcastle disease virus (NDV) and Sendai virus (SeV) showed low similarity (identity: 33–50%). It suggests that

henipavirus V proteins commonly interact with UBXN1, but other paramyxovirus V proteins might not have a similar interaction with it. UBXN1 belongs to the family of UBA–UBX-domain-containing

proteins (UBXNs). The UBXNs are considered to be the cofactors of an AAA ATPase, p97, which is involved in a large variety of cellular processes, including ubiquitin-dependent proteolysis,

the fusion of homotypic membranes, nuclear envelope reassembly, and cell-cycle progression44. UBXN1 has been shown to interact with p97 and polyubiquitin, which negatively regulates the

proteolysis of ubiquitylated proteins28,45. We confirmed the ubiquitylation of NiV V, but the expression levels of NiV V were unaffected by increased amounts of exogenous UBXN1 (data not

shown), suggesting that the proteolysis of NiV V is not regulated by UBXN1. The stabilization of UBXN1 by NiV V requires the UBX domain of UBXN1, and the UBX domain adopts the same

three-dimensional fold as ubiquitin46. Therefore, we speculate that the UBX domain is the determinant of the proteolytic rate of UBXN1, which is recognized by the proteasome system in a

similar way to ubiquitin, and that the V protein blocks its recognition, resulting in the stabilization of UBXN1. It has been reported that mice are resistant to intraperitoneal Henipavirus

infection, despite the expression of a functional viral entry receptor. However, the deletion of the type I IFN receptor makes them susceptible to this infection47. In cell lines and primary

endothelial cells, pretreatment with IFN also inhibited the replication of NiV14. These reports suggest the significance of the type I IFN system in controlling lethal Henipavirus

infections. A recombinant NiV lacking the V protein induced more IFNβ in primary human microvascular lung endothelial cells in a late step of infection than the wild-type virus48, suggesting

that the stabilization of UBXN1 by the V protein may contribute to the suppression of IFNβ induction in addition to its many reported IFN-suppressive functions. We believe that the

infection experiments using infectious NiV at biosafety level 4 would reveal the role of the potential ability of V protein to stabilize UBXN1 in the viral replication and pathogenesis in

future. METHODS CELLS HeLa, Huh-7, HEK293 and HEK293T cells were maintained in Dulbecco’s modified Eagle’s medium (Sigma) supplemented with 10% fetal calf serum under 5% CO2 at 37 °C.

Transfection was performed with Lipofectamine LTX (Invitrogen) according to the manufacturer’s instructions. The conditions of transfection for each experiment are not unified. PLASMIDS To

construct the expression vectors for viral proteins, pNiV(6+)49 were used as the polymerase chain reaction (PCR) templates. The expression vectors for untagged NiV P and V proteins were

constructed by inserting the PCR-amplified cDNA into pcDNA3.1(+) (Invitrogen). The expression vectors for myc-tagged NiV P and V were constructed by inserting the PCR-amplified cDNA into

pCMV-myc (Clontech). The deletion mutants of NiV V were constructed with inverted PCR using various combinations of primers, which were phosphorylated with T4 PNK (Toyobo). After

electrophoresis and gel extraction, the purified PCR products were self-ligated with T4 DNA ligase (Promega). The alanine-substituted mutants of NiV V were constructed with inverted PCR

using various combinations of primers containing the mutated nucleotide sequences. To clone the host genes for use as PCR templates, the total RNA was extracted from HeLa cells with ISOGEN

(Nippon Gene), and the cDNA was synthesized by reverse transcription with PrimeScript Reverse Transcriptase (Takara) and an oligo(dT) primer. The expression vector for hemagglutinin

(HA)-tagged UBXN1 was constructed by inserting the PCR-amplified cDNA into pCMV-HA (Clontech). The expression vectors for glutathione S-transferase (GST)-tagged UBXN1 were constructed by

inserting the PCR-amplified cDNA into pGEX-4T-2 (GE Healthcare). The deletion mutants of GST-tagged UBXN1 were constructed with inverted PCR, as described above. The expression vectors for

FLAG-tagged MDA5 and MAVS were constructed by inserting the PCR-amplified cDNA into pFLAG-CMV (Sigma). The expression vector for constitutively active RIG-I (RIG-IΔ) which lacks C-terminal

domain corresponding to amino acids 735–925 was constructed by inserting the PCR-amplified cDNA into pcDNA3.1(+). To construct the interferon β (IFNβ) reporter vector, the HeLa genome was

extracted with the phenol–chloroform method and used as the PCR template. The promoter region of the IFNB gene, corresponding to nucleotides −300 to +25, was amplified by PCR and inserted

into pGL3-Basic (Promega). The expression vector for EGFP (pEGFP-C1) was purchased from Clontech. The vector for the genome editing to deplete UBXN1 gene (px330-UBXN1) was constructed by

inserting the cDNA corresponding to 62,678,723–62,678,742 in human chromosome 11 (5′-GAGAAGGCTCTGGCCCTCAC-3′) into the downstream of U6 promoter in px330 vector50. PCR was performed with KOD

-Plus- Neo DNA polymerase (Toyobo), and the nucleotide sequences of all the constructed vectors were verified with DNA sequencing. IMMUNOPRECIPITATION ASSAY AND PROTEOMIC ANALYSIS At 48 h

posttransfection, the cells were washed twice with phosphate-buffered saline (PBS), and lysed in whole-cell extract buffer (50 mM Tris-HCl [pH 8.0], 280 mM NaCl, 0.5% NP-40, 0.2 mM EDTA, 2 

mM EGTA, 10% glycerol, 1 mM dithiothreitol, 1 mM sodium vanadate) supplemented with cOmplete Protease Inhibitor Cocktail (Roche). The cell debris was removed by centrifugation at 17 900 g

for 10 min, and the supernatant was collected. For the immunoprecipitation of myc-tagged NiV V and HA-tagged UBXN1, Protein G Sepharose (GE Healthcare) conjugated with equal amounts of

anti-myc mouse monoclonal antibody (Clontech), anti-HA mouse monoclonal antibody (Sigma), and mouse control IgG (R&D Systems) was used. For the immunoprecipitation of endogenous MAVS,

Protein A Sepharose (GE Healthcare) conjugated with anti-MAVS (CT) antibody (Millipore) was used. For the immunoprecipitation of endogenous UBXN1, Protein A Sepharose (GE Healthcare)

conjugated with anti-UBXN1 rabbit polyclonal antibody (Proteintech) was used. After incubation with the cell lysate at 4 °C for 1 h, the beads were washed three times with wash buffer (50 mM

Tris-HCl [pH 8.0], 300 mM NaCl, 0.5% NP-40 10% glycerol), and the precipitated proteins were eluted with elution buffer (0.1 M glycine-HCl [pH 2.0]). The immunoprecipitated proteins were

separated with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and detected with silver staining (Apro Science). The bands were excised from the polyacrylamide gel and

destained. After digestion with trypsin, the peptides were extracted with 5% (v/v) formic acid and acetonitrile, and analyzed with nanoscale liquid chromatography electrospray tandem mass

spectrometry. The detected peptides were analyzed with the Mascot search engine. GST PULL-DOWN ASSAY Escherichia coli strain BL21 was transformed with the vector expressing GST-tagged UBXN1,

and the expression of the recombinant protein was induced with 1 mM isopropyl β-D-1-thiogalactopyranoside at 30 °C for 3 h. After the bacterial cells were washed three times with PBS, they

were lysed and sonicated in lysis buffer (1% Triton X-100, 1% N-lauroylsarcosine in PBS). The cell debris was removed by centrifugation at 15 000 g for 15 min, and the supernatant was

incubated with Glutathione Sepharose 4B (GE Healthcare) at 4 °C for 1 h. After the beads were washed three times with wash buffer (described in the “immunoprecipitation assay” section), they

were incubated in the cell lysate at 4 °C for 1 h. The beads were washed three times, and the proteins were eluted with elution buffer (25 mM glutathione, 100 mM Tris-HCl [pH 8.9]). WESTERN

BLOTTING The proteins in the cell lysate and the eluted product were separated with SDS-PAGE and detected with western blotting. After the proteins were transferred to PVDF membrane, the

membrane was cut according to the bands of proteins marker (Precision Plus Protein™ Dual Color Standards, BIO-RAD) to probe multiple proteins. As the primary antibodies, we used anti-myc

rabbit polyclonal antibody (Sigma), anti-HA mouse monoclonal antibody (Sigma), anti-HA goat polyclonal antibody (Novus Biologicals), anti-FLAG mouse monoclonal antibody (Sigma), anti-UBXN1

rabbit polyclonal antibody (Millipore), anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mouse monoclonal antibody (Millipore), anti-GST mouse monoclonal antibody (Santa Cruz

Biotechnology), anti-MAVS rabbit polyclonal antibody (described in the “immunoprecipitation assay” section), anti-EGFP mouse monoclonal antibody (Clontech), and anti-NiV V rabbit polyclonal

antibody, which has been described previously51. As the secondary antibodies, horseradish-peroxidase-conjugated goat anti-mouse IgG antibody (Dako), anti-rabbit IgG antibody (Dako), or

anti-goat IgG antibody (Dako) were used. Chemiluminescence was detected with ECL Prime Western blotting Detection Reagent (GE Healthcare) and an ImageQuant LAS 4000 biomolecular imager (GE

Healthcare). CYCLOHEXIMIDE CHASE At 24 h posttransfection, the cells were trypsinized and split into five equal portions. After 24 h, the cells were treated with 100 µg/ml cycloheximide

(Sigma), and lysed as described in the “immunoprecipitation assay” section. GENOME EDITING The cells were transfected with px330-UBXN1 together with pQCXIP (Clontech), and were treated with

2 µg/ml puromycin (Sigma) for 9 days. After the cells were maintained in the medium without puromycin for 5 days, the depletion of UBXN1 was verified with western blotting analysis. INDIRECT

IMMUNOFLUORESCENCE ASSAY At 24 h posttransfection, the cells were fixed with 4% paraformaldehyde for 30 min. After the cells were washed three times with PBS, they were incubated in

blocking buffer (3% bovine serum albumin, 0.1% Triton X-100 in PBS) at room temperature for 30 min. For the staining of myc-tagged NiV V, UBXN1 and HA-tagged UBXN1, anti-myc rabbit

polyclonal antibody (Sigma), anti-myc mouse monoclonal antibody (Clontech), anti-UBXN1 rabbit polyclonal antibody (Millipore) and anti-HA mouse monoclonal antibody (Sigma) were incubated

with the cell in blocking buffer at 4 °C for over night. For control staining, rabbit serum was used. After the cells were washed three times with wash buffer (0.05% Tween 20 in PBS), they

were incubated with Alexa-Fluor-488-conjugated goat anti-rabbit antibody (Invitrogen), Alexa-Fluor-568-conjugated goat anti-mouse antibody (Invitrogen), and Hoechst 33342 (Cambrex) in

blocking buffer at room temperature for 1 h. After the cells were washed three times, their immunofluorescence was observed with an IX70 laser confocal microscope and the FluoView FV1000

system (Olympus). LUCIFERASE ASSAY For the MDA5 and RIG-I signaling assay, the cells were transfected with the IFNβ reporter vector and phRL-TK (Promega) as the internal control, together

with vectors expressing MDA5, RIG-IΔ, NiV V and UBXN1. At 24 h posttransfection, the cells were washed once with PBS and lysed in Passive Lysis Buffer (Promega). The luciferase activities in

the cell lysates were measured with the Dual Luciferase Assay System (Promega). QUANTITATIVE PCR ASSAY The template DNA fragment of the NiV minigenome RNA was described previously51. The

RNA was synthesized with the T7 RiboMax™ Express Large Scale RNA Production System (Promega) and purified according to the manufacturer’s instructions. After the quality of the synthesized

RNA was checked with formaldehyde gel electrophoresis, the RNA was transfected using Lipofectamine 2000 (Invitrogen). Total RNA was extracted from the cells with TRIzol Reagent (Invitrogen),

and was reverse-transcribed with PrimeScript Reverse Transcriptase (Takara) and an oligo(dT) primer. The quantitative PCR was performed with THUNDERBIRD® SYBR® qPCR Mix (Toyobo) and

specific primers for IFNβ (forward: 5′-CAGGAGAGCAATTTGGAGGA-3′; reverse: 5′-CTTTCGAAGCCTTTGCTCTG-3′) and β-actin (forward: 5′-TGGACTTCGAGCAAGAGATGG-3′; reverse: 5′-GGAAGGAAGGCTGGAAGAGTG-3′).

STATISTICAL ANALYSIS Quantitative data were presented as means ± standard deviations of triplicate samples. For the assessment of the normality, Kolmogorov-Smirnov test was used. The

statistical significance between two groups were examined by Student’s _t_ test after their homoscedasticity was assessed by F test. For the multiple comparison, one-way analysis of variance

(ANOVA) followed by Dunnett’s multiple comparison test was used. _P_ value of <0.05 was considered statistically significant. REFERENCES * Chua, K. B. _et al_. Nipah virus: a recently

emergent deadly paramyxovirus. _Science_ 288, 1432–1435 (2000). Article  ADS  CAS  PubMed  Google Scholar  * Eaton, B. T., Broder, C. C., Middleton, D. & Wang, L. F. Hendra and Nipah

viruses: different and dangerous. _Nat Rev Microbiol_ 4, 23–35, https://doi.org/10.1038/nrmicro1323 (2006). Article  CAS  PubMed  Google Scholar  * Arankalle, V. A. _et al_. Genomic

characterization of Nipah virus, West Bengal, India. _Emerg Infect Dis_ 17, 907–909, https://doi.org/10.3201/eid1705.100968 (2011). Article  PubMed  PubMed Central  Google Scholar  * Chadha,

M. S. _et al_. Nipah virus-associated encephalitis outbreak, Siliguri, India. _Emerg Infect Dis_ 12, 235–240, https://doi.org/10.3201/eid1202.051247 (2006). Article  PubMed  PubMed Central

  Google Scholar  * Ching, P. K. _et al_. Outbreak of henipavirus infection, Philippines, 2014. _Emerg Infect Dis_ 21, 328–331, https://doi.org/10.3201/eid2102.141433 (2015). Article  CAS 

PubMed  PubMed Central  Google Scholar  * Hossain, M. J. _et al_. Clinical presentation of nipah virus infection in Bangladesh. _Clin Infect Dis_ 46, 977–984, https://doi.org/10.1086/529147

(2008). Article  PubMed  Google Scholar  * Hsu, V. P. _et al_. Nipah virus encephalitis reemergence, Bangladesh. _Emerg Infect Dis_ 10, 2082–2087, https://doi.org/10.3201/eid1012.040701

(2004). Article  PubMed  PubMed Central  Google Scholar  * Pernet, O. _et al_. Evidence for henipavirus spillover into human populations in Africa. _Nat Commun_ 5, 5342,

https://doi.org/10.1038/ncomms6342 (2014). Article  PubMed  PubMed Central  Google Scholar  * Hayman, D. T. _et al_. Evidence of henipavirus infection in West African fruit bats. _PLoS One_

3, e2739, https://doi.org/10.1371/journal.pone.0002739 (2008). Article  ADS  PubMed  PubMed Central  Google Scholar  * Iehle, C. _et al_. Henipavirus and Tioman virus antibodies in

pteropodid bats, Madagascar. _Emerg Infect Dis_ 13, 159–161, https://doi.org/10.3201/eid1301.060791 (2007). Article  PubMed  PubMed Central  Google Scholar  * Li, Y. _et al_. Antibodies to

Nipah or Nipah-like viruses in bats, China. _Emerg Infect Dis_ 14, 1974–1976, https://doi.org/10.3201/eid1412.080359 (2008). Article  ADS  PubMed  PubMed Central  Google Scholar  * Harcourt,

B. H. _et al_. Molecular characterization of Nipah virus, a newly emergent paramyxovirus. _Virology_ 271, 334–349, https://doi.org/10.1006/viro.2000.0340 (2000). Article  CAS  PubMed 

Google Scholar  * Yoneda, M. _et al_. The nonstructural proteins of Nipah virus play a key role in pathogenicity in experimentally infected animals. _PLoS One_ 5, e12709,

https://doi.org/10.1371/journal.pone.0012709 (2010). Article  ADS  PubMed  PubMed Central  Google Scholar  * Satterfield, B. A. _et al_. The immunomodulating V and W proteins of Nipah virus

determine disease course. _Nat Commun_ 6, 7483, https://doi.org/10.1038/ncomms8483 (2015). Article  CAS  PubMed  PubMed Central  Google Scholar  * Satterfield, B. A. _et al_. Nipah Virus C

and W Proteins Contribute to Respiratory Disease in Ferrets. _J Virol_ 90, 6326–6343, https://doi.org/10.1128/jvi.00215-16 (2016). Article  CAS  PubMed  PubMed Central  Google Scholar  *

Mathieu, C. _et al_. Nonstructural Nipah virus C protein regulates both the early host proinflammatory response and viral virulence. _J Virol_ 86, 10766–10775,

https://doi.org/10.1128/jvi.01203-12 (2012). Article  CAS  PubMed  PubMed Central  Google Scholar  * Marsh, G. A. _et al_. Cedar virus: a novel Henipavirus isolated from Australian bats.

_PLoS Pathog_ 8, e1002836, https://doi.org/10.1371/journal.ppat.1002836 (2012). Article  CAS  PubMed  PubMed Central  Google Scholar  * Andrejeva, J. _et al_. The V proteins of

paramyxoviruses bind the IFN-inducible RNA helicase, mda-5, and inhibit its activation of the IFN-beta promoter. _Proc Natl Acad Sci USA_ 101, 17264–17269,

https://doi.org/10.1073/pnas.0407639101 (2004). Article  ADS  CAS  PubMed  PubMed Central  Google Scholar  * Childs, K. _et al_. mda-5, but not RIG-I, is a common target for paramyxovirus V

proteins. _Virology_ 359, 190–200, https://doi.org/10.1016/j.virol.2006.09.023 (2007). Article  CAS  PubMed  Google Scholar  * Davis, M. E. _et al_. Antagonism of the phosphatase PP1 by the

measles virus V protein is required for innate immune escape of MDA5. _Cell Host Microbe_ 16, 19–30, https://doi.org/10.1016/j.chom.2014.06.007 (2014). Article  CAS  PubMed  PubMed Central 

Google Scholar  * Childs, K., Randall, R. & Goodbourn, S. Paramyxovirus V proteins interact with the RNA Helicase LGP2 to inhibit RIG-I-dependent interferon induction. _J Virol_ 86,

3411–3421, https://doi.org/10.1128/jvi.06405-11 (2012). Article  CAS  PubMed  PubMed Central  Google Scholar  * Kitagawa, Y. _et al_. A tryptophan-rich motif in the human parainfluenza virus

type 2 V protein is critical for the blockade of toll-like receptor 7 (TLR7)- and TLR9-dependent signaling. _J Virol_ 85, 4606–4611, https://doi.org/10.1128/jvi.02012-10 (2011). Article 

CAS  PubMed  PubMed Central  Google Scholar  * Shaw, M. L., Cardenas, W. B., Zamarin, D., Palese, P. & Basler, C. F. Nuclear localization of the Nipah virus W protein allows for

inhibition of both virus- and toll-like receptor 3-triggered signaling pathways. _J Virol_ 79, 6078–6088, https://doi.org/10.1128/jvi.79.10.6078-6088.2005 (2005). Article  CAS  PubMed 

PubMed Central  Google Scholar  * Rodriguez, J. J., Parisien, J. P. & Horvath, C. M. Nipah virus V protein evades alpha and gamma interferons by preventing STAT1 and STAT2 activation and

nuclear accumulation. _J Virol_ 76, 11476–11483 (2002). Article  CAS  PubMed  PubMed Central  Google Scholar  * Rodriguez, J. J., Wang, L. F. & Horvath, C. M. Hendra virus V protein

inhibits interferon signaling by preventing STAT1 and STAT2 nuclear accumulation. _J Virol_ 77, 11842–11845 (2003). Article  CAS  PubMed  PubMed Central  Google Scholar  * Sleeman, K. _et

al_. The C, V and W proteins of Nipah virus inhibit minigenome replication. _J Gen Virol_ 89, 1300–1308, https://doi.org/10.1099/vir.0.83582-0 (2008). Article  CAS  PubMed  Google Scholar  *

McNeill, H., Knebel, A., Arthur, J. S., Cuenda, A. & Cohen, P. A novel UBA and UBX domain protein that binds polyubiquitin and VCP and is a substrate for SAPKs. _Biochem J_ 384,

391–400, https://doi.org/10.1042/bj20041498 (2004). Article  CAS  PubMed  PubMed Central  Google Scholar  * Ishibashi, T. _et al_. A novel protein specifically interacting with Homer2

regulates ubiquitin-proteasome systems. _J Biochem_ 137, 617–623, https://doi.org/10.1093/jb/mvi074 (2005). Article  CAS  PubMed  Google Scholar  * Ramachandran, A. & Horvath, C. M.

Dissociation of paramyxovirus interferon evasion activities: universal and virus-specific requirements for conserved V protein amino acids in MDA5 interference. _J Virol_ 84, 11152–11163,

https://doi.org/10.1128/jvi.01375-10 (2010). Article  CAS  PubMed  PubMed Central  Google Scholar  * Wang, P. _et al_. UBXN1 interferes with Rig-I-like receptor-mediated antiviral immune

response by targeting MAVS. _Cell Rep_ 3, 1057–1070, https://doi.org/10.1016/j.celrep.2013.02.027 (2013). Article  CAS  PubMed  Google Scholar  * Bowie, A. G. & Unterholzner, L. Viral

evasion and subversion of pattern-recognition receptor signalling. _Nat Rev Immunol_ 8, 911–922, https://doi.org/10.1038/nri2436 (2008). Article  CAS  PubMed  Google Scholar  * Hou, F. _et

al_. MAVS forms functional prion-like aggregates to activate and propagate antiviral innate immune response. _Cell_ 146, 448–461, https://doi.org/10.1016/j.cell.2011.06.041 (2011). Article 

CAS  PubMed  PubMed Central  Google Scholar  * Shaw, M. L., Garcia-Sastre, A., Palese, P. & Basler, C. F. Nipah virus V and W proteins have a common STAT1-binding domain yet inhibit

STAT1 activation from the cytoplasmic and nuclear compartments, respectively. _J Virol_ 78, 5633–5641, https://doi.org/10.1128/jvi.78.11.5633-5641.2004 (2004). Article  CAS  PubMed  PubMed

Central  Google Scholar  * Rodriguez, K. R. & Horvath, C. M. Paramyxovirus V protein interaction with the antiviral sensor LGP2 disrupts MDA5 signaling enhancement but is not relevant to

LGP2-mediated RLR signaling inhibition. _J Virol_ 88, 8180–8188, https://doi.org/10.1128/jvi.00737-14 (2014). Article  PubMed  PubMed Central  Google Scholar  * Irie, T., Kiyotani, K.,

Igarashi, T., Yoshida, A. & Sakaguchi, T. Inhibition of interferon regulatory factor 3 activation by paramyxovirus V protein. _J Virol_ 86, 7136–7145,

https://doi.org/10.1128/jvi.06705-11 (2012). Article  CAS  PubMed  PubMed Central  Google Scholar  * Wu-Baer, F., Ludwig, T. & Baer, R. The UBXN1 protein associates with

autoubiquitinated forms of the BRCA1 tumor suppressor and inhibits its enzymatic function. _Mol Cell Biol_ 30, 2787–2798, https://doi.org/10.1128/mcb.01056-09 (2010). Article  CAS  PubMed 

PubMed Central  Google Scholar  * Kubota, T., Yokosawa, N., Yokota, S. & Fujii, N. C terminal CYS-RICH region of mumps virus structural V protein correlates with block of interferon

alpha and gamma signal transduction pathway through decrease of STAT 1-alpha. _Biochem Biophys Res Commun_ 283, 255–259, https://doi.org/10.1006/bbrc.2001.4764 (2001). Article  CAS  PubMed 

Google Scholar  * Palosaari, H., Parisien, J. P., Rodriguez, J. J., Ulane, C. M. & Horvath, C. M. STAT protein interference and suppression of cytokine signal transduction by measles

virus V protein. _J Virol_ 77, 7635–7644 (2003). Article  CAS  PubMed  PubMed Central  Google Scholar  * Parisien, J. P. _et al_. The V protein of human parainfluenza virus 2 antagonizes

type I interferon responses by destabilizing signal transducer and activator of transcription 2. _Virology_ 283, 230–239, https://doi.org/10.1006/viro.2001.0856 (2001). Article  CAS  PubMed

  Google Scholar  * Parisien, J. P., Lau, J. F., Rodriguez, J. J., Ulane, C. M. & Horvath, C. M. Selective STAT protein degradation induced by paramyxoviruses requires both STAT1 and

STAT2 but is independent of alpha/beta interferon signal transduction. _J Virol_ 76, 4190–4198 (2002). Article  CAS  PubMed  PubMed Central  Google Scholar  * Takeuchi, K., Kadota, S. I.,

Takeda, M., Miyajima, N. & Nagata, K. Measles virus V protein blocks interferon (IFN)-alpha/beta but not IFN-gamma signaling by inhibiting STAT1 and STAT2 phosphorylation. _FEBS Lett_

545, 177–182 (2003). Article  CAS  PubMed  Google Scholar  * Ulane, C. M. & Horvath, C. M. Paramyxoviruses SV5 and HPIV2 assemble STAT protein ubiquitin ligase complexes from cellular

components. _Virology_ 304, 160–166 (2002). Article  CAS  PubMed  Google Scholar  * Yokosawa, N., Kubota, T. & Fujii, N. Poor induction of interferon-induced 2′,5′-oligoadenylate

synthetase (2–5 AS) in cells persistently infected with mumps virus is caused by decrease of STAT-1 alpha. _Arch Virol_ 143, 1985–1992 (1998). Article  CAS  PubMed  Google Scholar  *

Schuberth, C. & Buchberger, A. UBX domain proteins: major regulators of the AAA ATPase Cdc48/p97. _Cell Mol Life Sci_ 65, 2360–2371, https://doi.org/10.1007/s00018-008-8072-8 (2008).

Article  CAS  PubMed  Google Scholar  * LaLonde, D. P. & Bretscher, A. The UBX protein SAKS1 negatively regulates endoplasmic reticulum-associated degradation and p97-dependent

degradation. _J Biol Chem_ 286, 4892–4901, https://doi.org/10.1074/jbc.M110.158030 (2011). Article  CAS  PubMed  Google Scholar  * Buchberger, A., Howard, M. J., Proctor, M. & Bycroft,

M. The UBX domain: a widespread ubiquitin-like module. _J Mol Biol_ 307, 17–24, https://doi.org/10.1006/jmbi.2000.4462 (2001). Article  CAS  PubMed  Google Scholar  * Dhondt, K. P. _et al_.

Type I interferon signaling protects mice from lethal henipavirus infection. _J Infect Dis_ 207, 142–151, https://doi.org/10.1093/infdis/jis653 (2013). Article  CAS  PubMed  Google Scholar 

* Lo, M. K. _et al_. Distinct and overlapping roles of Nipah virus P gene products in modulating the human endothelial cell antiviral response. _PLoS One_ 7, e47790,

https://doi.org/10.1371/journal.pone.0047790 (2012). Article  ADS  CAS  PubMed  PubMed Central  Google Scholar  * Yoneda, M. _et al_. Establishment of a Nipah virus rescue system. _Proc Natl

Acad Sci USA_ 103, 16508–16513, https://doi.org/10.1073/pnas.0606972103 (2006). Article  ADS  CAS  PubMed  PubMed Central  Google Scholar  * Cong, L. _et al_. Multiplex genome engineering

using CRISPR/Cas systems. _Science_ 339, 819–823, https://doi.org/10.1126/science.1231143 (2013). Article  ADS  CAS  PubMed  PubMed Central  Google Scholar  * Omi-Furutani, M., Yoneda, M.,

Fujita, K., Ikeda, F. & Kai, C. Novel phosphoprotein-interacting region in Nipah virus nucleocapsid protein and its involvement in viral replication. _J Virol_ 84, 9793–9799,

https://doi.org/10.1128/jvi.00339-10 (2010). Article  CAS  PubMed  PubMed Central  Google Scholar  Download references ACKNOWLEDGEMENTS This work was supported by Grants-in-Aid for

Scientific Research (KAKENHI) from the Japan Society for the Promotion of Science (JSPS). The mass spectrometry analysis was conducted by the Laboratory Center for Proteomics Research

(Institution of Medical Science, The University of Tokyo). AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Laboratory Animal Research Center and International Research Center for Infectious

Diseases, Institute of Medical Science, The University of Tokyo, Tokyo, Japan Shotaro Uchida, Ryo Horie, Hiroki Sato, Chieko Kai & Misako Yoneda Authors * Shotaro Uchida View author

publications You can also search for this author inPubMed Google Scholar * Ryo Horie View author publications You can also search for this author inPubMed Google Scholar * Hiroki Sato View

author publications You can also search for this author inPubMed Google Scholar * Chieko Kai View author publications You can also search for this author inPubMed Google Scholar * Misako

Yoneda View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS S.U. have performed all experiments, and prepared the manuscript. R.H. have assisted

the experiments. H.S. and C.K. have provided materials and intellectual support. M.Y. have directed the entire study and manuscript preparation. All authors discussed the results and

contributed to revision of the manuscript. CORRESPONDING AUTHOR Correspondence to Misako Yoneda. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing interests.

ADDITIONAL INFORMATION PUBLISHER'S NOTE: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. ELECTRONIC SUPPLEMENTARY

MATERIAL SUPPLEMENTARY INFORMATION RIGHTS AND PERMISSIONS OPEN ACCESS This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing,

adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons

license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a

credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted

use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Reprints and permissions ABOUT

THIS ARTICLE CITE THIS ARTICLE Uchida, S., Horie, R., Sato, H. _et al._ Possible role of the Nipah virus V protein in the regulation of the interferon beta induction by interacting with UBX

domain-containing protein1. _Sci Rep_ 8, 7682 (2018). https://doi.org/10.1038/s41598-018-25815-9 Download citation * Received: 14 August 2017 * Accepted: 17 April 2018 * Published: 16 May

2018 * DOI: https://doi.org/10.1038/s41598-018-25815-9 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a shareable

link is not currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative