Rnase h hydrolysis of the 5′ terminus of the avian sarcoma virus genome during reverse transcription

ABSTRACT NUCLEOTIDE sequence analyses of the ends of the avian retro-virus genome 1–6 have confirmed predictions of the terminally redundant nature of the viral RNA7,8. These studies have

also provided the basis for several models of retrovirus proviral DNA synthesis, all of which have implicated the RNase H activity associated with viral reverse transcriptase in a functional

role in the continued and uninterrupted DNA transcription of the viral RNA genome1–8. According to these models DNA synthesis initiates on the tRNAtrp primer molecule located close to the

5′ end of the viral genome9,10 and transcription proceeds to the terminus. Presumably, at this time a suitable substrate is available for the retrovirus RNase H, which is a processive

exoribonuclease requiring an unblocked terminus of the RNA moiety of RNA:DNA hybrids for activity11–13. Release of the terminally repeated nucleotides from the hybrid region of the viral

genome would create ‘sticky ends’ with the tRNAtrp-initiated DNA at the 5′ end capable of hybridising to the terminally redundant RNA genomic sequences at the 3′ end of the same or a second

35S RNA subunit. This latter reaction would facilitate uninterrupted transcription from the 5′ to the 3′ end of the viral genome resulting in genome-length DNA transcripts15–20. We have

recently obtained evidence indicating that DNA transcripts much longer than the distance between the tRNAtrp primer molecule and the 5′ end of the viral genome and containing nucleotide

sequences representing the 3′ region of the viral genome can be synthesised by the reverse transcriptase _in vitro_21. Thus, it seems that DNA synthesis initiated at the 5′ end of the viral

genome continues at the 3′ end in enzymatic reactions _in vitro_. If the avian sarcoma virus (ASV) reverse transcriptase-associated RNase H activity is required for the continued

transcription of the viral genome at the 3′ end as proposed in above models, then release of 5′ terminally-located ribonucleotides should be apparent during DNA synthesis _in vitro_. We

report here that ribonucleotides are indeed released from the viral RNA genome during reverse transcription, and that hydrolysis occurs at a specific site near the 5′ terminus. These studies

exemplify the first demonstration of RNase H hydrolysis occurring during reverse transcription of the retro-virus RNA genome _in vitro_ and implicate functional role for this activity

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December 2024 TUT4/7-MEDIATED URIDYLATION OF A CORONAVIRUS SUBGENOMIC RNAS DELAYS VIRAL REPLICATION Article Open access 21 April 2023 THE HOST RNA POLYMERASE II C-TERMINAL DOMAIN IS THE

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_Nature_ 257, 618–620 (1975). Article  ADS  CAS  PubMed  Google Scholar  Download references AUTHOR INFORMATION Author notes * MARC S. COLLETT Present address: Department of Pathology,

University of Colorado, Medical Center, Denver, Colorado, 802620 AUTHORS AND AFFILIATIONS * Department of Microbiology, University of Virginia Medical School, Charlottesville, Virginia,

22901 MARC S. COLLETT, PETER DIERKS & J. THOMAS PARSONS * Department of Microbiology, University of Minnesota Medical School, Minneapolis, Minnesota, 55455 ANTHONY J. FARAS Authors *

MARC S. COLLETT View author publications You can also search for this author inPubMed Google Scholar * PETER DIERKS View author publications You can also search for this author inPubMed 

Google Scholar * J. THOMAS PARSONS View author publications You can also search for this author inPubMed Google Scholar * ANTHONY J. FARAS View author publications You can also search for

this author inPubMed Google Scholar RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE COLLETT, M., DIERKS, P., PARSONS, J. _et al._ RNase H hydrolysis of

the 5′ terminus of the avian sarcoma virus genome during reverse transcription. _Nature_ 272, 181–184 (1978). https://doi.org/10.1038/272181a0 Download citation * Received: 25 August 1977 *

Accepted: 14 November 1977 * Issue Date: 09 March 1978 * DOI: https://doi.org/10.1038/272181a0 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content:

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