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Cis-Allosteric Regulation of HIV-1 Reverse Transcriptase by Integrase.
Masuda, Takao; Kotani, Osamu; Yokoyama, Masaru; Abe, Yuya; Kawai, Gota; Sato, Hironori.
Afiliação
  • Masuda T; Department of Immunotherapeutics, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Yushima, 1-5-45 Bunkyo-ku, Tokyo 113-8519, Japan.
  • Kotani O; Laboratory of Viral Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases, Gakuen, 4-7-1, Musashimurayama-shi, Tokyo 208-0011, Japan.
  • Yokoyama M; Laboratory of Viral Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases, Gakuen, 4-7-1, Musashimurayama-shi, Tokyo 208-0011, Japan.
  • Abe Y; Department of Immunotherapeutics, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Yushima, 1-5-45 Bunkyo-ku, Tokyo 113-8519, Japan.
  • Kawai G; Department of Life Science, Faculty of Advanced Engineering, Chiba Institute of Technology, 2-17-1 Tsudanuma, Narashino-shi, Chiba 275-0016, Japan.
  • Sato H; Laboratory of Viral Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases, Gakuen, 4-7-1, Musashimurayama-shi, Tokyo 208-0011, Japan.
Viruses ; 15(1)2022 12 21.
Article em En | MEDLINE | ID: mdl-36680070
Reverse transcriptase (RT) and integrase (IN) are encoded tandemly in the pol genes of retroviruses. We reported recently that HIV-1 RT and IN need to be supplied as the pol precursor intermediates, in which RT and IN are in fusion form (RTIN) to exert efficient reverse transcription in the context of HIV-1 replication. The mechanism underlying RTIN's effect, however, remains to be elucidated. In this study, we examined the effect of IN fusion on RT during reverse transcription by an in vitro cell-free assay, using recombinant HIV-1 RTIN (rRTIN). We found that, compared to recombinant RT (rRT), rRTIN generated significantly higher cDNAs under physiological concentrations of dNTPs (less than 10 µM), suggesting increased affinity of RTIN to dNTPs. Importantly, the cleavage of RTIN with HIV-1 protease reduced cDNA levels at a low dose of dNTPs. Similarly, sensitivities against RT inhibitors were significantly altered in RTIN form. Finally, analysis of molecular dynamics simulations of RT and RTIN suggested that IN can influence the structural dynamics of the RT active center and the inhibitor binding pockets in cis. Thus, we demonstrated, for the first time, the cis-allosteric regulatory roles of IN in RT structure and enzymatic activity.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Integrase de HIV / Transcriptase Reversa do HIV Idioma: En Revista: Viruses Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Japão

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Integrase de HIV / Transcriptase Reversa do HIV Idioma: En Revista: Viruses Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Japão