| RRC ID |
77417
|
| 著者 |
Masuda T, Sato Y, Huang YL, Koi S, Takahata T, Hasegawa A, Kawai G, Kannagi M.
|
| タイトル |
Fate of HIV-1 cDNA intermediates during reverse transcription is dictated by transcription initiation site of virus genomic RNA.
|
| ジャーナル |
Sci Rep
|
| Abstract |
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.
|
| 巻・号 |
5
|
| ページ |
17680
|
| 公開日 |
2015-12-3
|
| DOI |
10.1038/srep17680
|
| PII |
srep17680
|
| PMID |
26631448
|
| PMC |
PMC4668388
|
| MeSH |
DNA, Complementary / genetics*
HEK293 Cells / virology
HIV-1 / genetics*
HIV-1 / pathogenicity
Humans
Nucleocapsid Proteins / genetics
Nucleocapsid Proteins / metabolism
RNA, Viral / genetics*
Reverse Transcription*
Transcription Initiation Site*
|
| IF |
3.998
|
| リソース情報 |
| 遺伝子材料 |
CSII-CMV-MCS (RDB04377) |
