title: Topoisomerase I and ATP activate cDNA synthesis of human immunodeficiency virus type 1.
abstract: Replication of human immunodeficiency virus type 1 (HIV-1) is regulated at reverse transcription. Cellular topoisomerase I has been reported to be carried into HIV-1 virions and enhance cDNA synthesis in vitro, suggesting that topoisomerase I expressed in virus producer cells regulates reverse transcription. Here, by employing both indicator cell assay and endogenous reverse transcription (ERT) assay, we show that topoisomerase I and adenosine triphosphate (ATP) enhanced cDNA synthesis of HIV-1. In addition, topoisomerase I mutants, R488A and K532A, lacking enzymatic activity, attenuated the efficiency of cDNA synthesis and resulted in inhibition of the infectivity of HIV-1, suggesting that the activity of topoisomerase I lacking in these mutants is indispensable for the cDNA synthesis in the HIV-1 replication process. Furthermore, ATP could dissociate topoisomerase I from the topoisomerase I-RNA complex and enhance cDNA synthesis in vitro. These findings suggest that cellular topoisomerase I and ATP play a pivotal role in the synthesis of cDNA of HIV-1.
authors.affiliation: Department of Pathology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan. htakahas@nih.go.jp
authors.forename: Hidehiro
authors.initials: H
authors.lastname: Takahashi
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Sawa
authors.forename: Hideki
authors.initials: H
authors.lastname: Hasegawa
authors.forename: Yuko
authors.initials: Y
authors.lastname: Shoya
authors.forename: Tetsutaro
authors.initials: T
authors.lastname: Sata
authors.forename: William W
authors.initials: WW
authors.lastname: Hall
authors.forename: Kazuo
authors.initials: K
authors.lastname: Nagashima
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kurata
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism* / Q000494:pharmacology; D019943:Amino Acid Substitution / Q000235:genetics; D004264:DNA Topoisomerases, Type I / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D018076:DNA, Complementary / Q000096:biosynthesis*; D017930:Genes, Reporter; D054303:HIV Reverse Transcriptase / Q000378:metabolism; D015497:HIV-1 / Q000254:growth & development / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D046911:Macromolecular Substances; D012367:RNA, Viral / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects / Q000502:physiology*
chemical_list: D018076:DNA, Complementary; D046911:Macromolecular Substances; D012367:RNA, Viral; C022571:adenosine 5'-O-(3-thiotriphosphate); D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D054303:HIV Reverse Transcriptase; D004264:DNA Topoisomerases, Type I