Cargando…
Fate of HIV-1 cDNA intermediates during reverse transcription is dictated by transcription initiation site of virus genomic RNA
Retroviral reverse transcription is accomplished by sequential strand-transfers of partial cDNA intermediates copied from viral genomic RNA. Here, we revealed an unprecedented role of 5′-end guanosine (G) of HIV-1 genomic RNA for reverse transcription. Based on current consensus for HIV-1 transcript...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4668388/ https://www.ncbi.nlm.nih.gov/pubmed/26631448 http://dx.doi.org/10.1038/srep17680 |
_version_ | 1782403983128657920 |
---|---|
author | Masuda, Takao Sato, Yoko Huang, Yu-Lun Koi, Satoshi Takahata, Tatsuro Hasegawa, Atsuhiko Kawai, Gota Kannagi, Mari |
author_facet | Masuda, Takao Sato, Yoko Huang, Yu-Lun Koi, Satoshi Takahata, Tatsuro Hasegawa, Atsuhiko Kawai, Gota Kannagi, Mari |
author_sort | Masuda, Takao |
collection | PubMed |
description | Retroviral reverse transcription is accomplished by sequential strand-transfers of partial cDNA intermediates copied from viral genomic RNA. Here, we revealed an unprecedented role of 5′-end guanosine (G) of HIV-1 genomic RNA for reverse transcription. Based on current consensus for HIV-1 transcription initiation site, HIV-1 transcripts possess a single G at 5′-ends (G1-form). However, we found that HIV-1 transcripts with additional Gs at 5′-ends (G2- and G3-forms) were abundantly expressed in infected cells by using alternative transcription initiation sites. The G2- and G3-forms were also detected in the virus particle, although the G1-form predominated. To address biological impact of the 5′-G number, we generated HIV clone DNA to express the G1-form exclusively by deleting the alternative initiation sites. Virus produced from the clone showed significantly higher strand-transfer of minus strong-stop cDNA (-sscDNA). The in vitro assay using synthetic HIV-1 RNAs revealed that the abortive forms of -sscDNA were abundantly generated from the G3-form RNA, but dramatically reduced from the G1-form. Moreover, the strand-transfer of -sscDNA from the G1-form was prominently stimulated by HIV-1 nucleocapsid. Taken together, our results demonstrated that the 5′-G number that corresponds to HIV-1 transcription initiation site was critical for successful strand-transfer of -sscDNA during reverse transcription. |
format | Online Article Text |
id | pubmed-4668388 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-46683882015-12-09 Fate of HIV-1 cDNA intermediates during reverse transcription is dictated by transcription initiation site of virus genomic RNA Masuda, Takao Sato, Yoko Huang, Yu-Lun Koi, Satoshi Takahata, Tatsuro Hasegawa, Atsuhiko Kawai, Gota Kannagi, Mari Sci Rep Article Retroviral reverse transcription is accomplished by sequential strand-transfers of partial cDNA intermediates copied from viral genomic RNA. Here, we revealed an unprecedented role of 5′-end guanosine (G) of HIV-1 genomic RNA for reverse transcription. Based on current consensus for HIV-1 transcription initiation site, HIV-1 transcripts possess a single G at 5′-ends (G1-form). However, we found that HIV-1 transcripts with additional Gs at 5′-ends (G2- and G3-forms) were abundantly expressed in infected cells by using alternative transcription initiation sites. The G2- and G3-forms were also detected in the virus particle, although the G1-form predominated. To address biological impact of the 5′-G number, we generated HIV clone DNA to express the G1-form exclusively by deleting the alternative initiation sites. Virus produced from the clone showed significantly higher strand-transfer of minus strong-stop cDNA (-sscDNA). The in vitro assay using synthetic HIV-1 RNAs revealed that the abortive forms of -sscDNA were abundantly generated from the G3-form RNA, but dramatically reduced from the G1-form. Moreover, the strand-transfer of -sscDNA from the G1-form was prominently stimulated by HIV-1 nucleocapsid. Taken together, our results demonstrated that the 5′-G number that corresponds to HIV-1 transcription initiation site was critical for successful strand-transfer of -sscDNA during reverse transcription. Nature Publishing Group 2015-12-03 /pmc/articles/PMC4668388/ /pubmed/26631448 http://dx.doi.org/10.1038/srep17680 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Masuda, Takao Sato, Yoko Huang, Yu-Lun Koi, Satoshi Takahata, Tatsuro Hasegawa, Atsuhiko Kawai, Gota Kannagi, Mari Fate of HIV-1 cDNA intermediates during reverse transcription is dictated by transcription initiation site of virus genomic RNA |
title | Fate of HIV-1 cDNA intermediates during reverse transcription is dictated by transcription initiation site of virus genomic RNA |
title_full | Fate of HIV-1 cDNA intermediates during reverse transcription is dictated by transcription initiation site of virus genomic RNA |
title_fullStr | Fate of HIV-1 cDNA intermediates during reverse transcription is dictated by transcription initiation site of virus genomic RNA |
title_full_unstemmed | Fate of HIV-1 cDNA intermediates during reverse transcription is dictated by transcription initiation site of virus genomic RNA |
title_short | Fate of HIV-1 cDNA intermediates during reverse transcription is dictated by transcription initiation site of virus genomic RNA |
title_sort | fate of hiv-1 cdna intermediates during reverse transcription is dictated by transcription initiation site of virus genomic rna |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4668388/ https://www.ncbi.nlm.nih.gov/pubmed/26631448 http://dx.doi.org/10.1038/srep17680 |
work_keys_str_mv | AT masudatakao fateofhiv1cdnaintermediatesduringreversetranscriptionisdictatedbytranscriptioninitiationsiteofvirusgenomicrna AT satoyoko fateofhiv1cdnaintermediatesduringreversetranscriptionisdictatedbytranscriptioninitiationsiteofvirusgenomicrna AT huangyulun fateofhiv1cdnaintermediatesduringreversetranscriptionisdictatedbytranscriptioninitiationsiteofvirusgenomicrna AT koisatoshi fateofhiv1cdnaintermediatesduringreversetranscriptionisdictatedbytranscriptioninitiationsiteofvirusgenomicrna AT takahatatatsuro fateofhiv1cdnaintermediatesduringreversetranscriptionisdictatedbytranscriptioninitiationsiteofvirusgenomicrna AT hasegawaatsuhiko fateofhiv1cdnaintermediatesduringreversetranscriptionisdictatedbytranscriptioninitiationsiteofvirusgenomicrna AT kawaigota fateofhiv1cdnaintermediatesduringreversetranscriptionisdictatedbytranscriptioninitiationsiteofvirusgenomicrna AT kannagimari fateofhiv1cdnaintermediatesduringreversetranscriptionisdictatedbytranscriptioninitiationsiteofvirusgenomicrna |