Počet záznamů: 1
Mechanism of polypurine tract primer generation by HIV-1 reverse transcriptase
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SYSNO ASEP 0486050 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Mechanism of polypurine tract primer generation by HIV-1 reverse transcriptase Tvůrce(i) Figiel, M. (PL)
Krepl, Miroslav (BFU-R) ORCID
Park, S. (US)
Poznanski, J. (PL)
Skowronek, K. (PL)
Golab, A. (PL)
Ha, T. (US)
Šponer, Jiří (BFU-R) RID, ORCID
Nowotny, M. (PL)Celkový počet autorů 9 Zdroj.dok. Journal of Biological Chemistry. - : Elsevier - ISSN 0021-9258
Roč. 293, č. 1 (2018), s. 191-202Poč.str. 12 s. Forma vydání Tištěná - P Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova human-immunodeficiency-virus ; strand dna-synthesis ; retroviral rnases-h ; rna/dna hybrid Vědní obor RIV CE - Biochemie Obor OECD Biochemistry and molecular biology CEP GBP305/12/G034 GA ČR - Grantová agentura ČR Institucionální podpora BFU-R - RVO:68081707 UT WOS 000419453200017 DOI 10.1074/jbc.M117.798256 Anotace HIV-1 reverse transcriptase (RT) possesses both DNA polymerase activity and RNase H activity that act in concert to convert single-stranded RNA of the viral genome to double-stranded DNA that is then integrated into the DNA of the infected cell. Reverse transcriptase-catalyzed reverse transcription critically relies on the proper generation of a polypurine tract (PPT) primer. However, the mechanism of PPT primer generation and the features of the PPT sequence that are critical for its recognition by HIV-1 RT remain unclear. Here, we used a chemical cross-linking method together with molecular dynamics simulations and single-molecule assays to study the mechanism of PPT primer generation. We found that the PPT was specifically and properly recognized within covalently tethered HIV-1 RT-nucleic acid complexes. These findings indicated that recognition of the PPT occurs within a stable catalytic complex after its formation. We found that this unique recognition is based on two complementary elements that rely on the PPT sequence: RNase H sequence preference and incompatibility of the poly(rA/dT) tract of the PPT with the nucleic acid conformation that is required for RNase H cleavage. The latter results from rigidity of the poly(rA/dT) tract and leads to base-pair slip-page of this sequence upon deformation into a catalytically relevant geometry. In summary, our results reveal an unexpected mechanism of PPT primer generation based on specific dynamic properties of the poly(rA/dT) segment and help advance our understanding of the mechanisms in viral RNA reverse transcription. Pracoviště Biofyzikální ústav Kontakt Jana Poláková, polakova@ibp.cz, Tel.: 541 517 244 Rok sběru 2018
Počet záznamů: 1