title: High fidelity simian immunodeficiency virus reverse transcriptase mutants have impaired replication in vitro and in vivo.
abstract: The low fidelity of HIV replication facilitates immune and drug escape. Some reverse transcriptase (RT) inhibitor drug-resistance mutations increase RT fidelity in biochemical assays but their effect during viral replication is unclear. We investigated the effect of RT mutations K65R, Q151N and V148I on SIV replication and fidelity in vitro, along with SIV replication in pigtailed macaques. SIVmac239-K65R and SIVmac239-V148I viruses had reduced replication capacity compared to wild-type SIVmac239. Direct virus competition assays demonstrated a rank order of wild-type>K65R>V148I mutants in terms of viral fitness. In single round in vitro-replication assays, SIVmac239-K65R demonstrated significantly higher fidelity than wild-type, and rapidly reverted to wild-type following infection of macaques. In contrast, SIVmac239-Q151N was replication incompetent in vitro and in pigtailed macaques. Thus, we showed that RT mutants, and specifically the common K65R drug-resistance mutation, had impaired replication capacity and higher fidelity. These results have implications for the pathogenesis of drug-resistant HIV.
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, The Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
authors.forename: Sarah B
authors.initials: SB
authors.lastname: Lloyd
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, The Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
authors.forename: Marit
authors.initials: M
authors.lastname: Lichtfuss
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, The Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
authors.forename: Thakshila H
authors.initials: TH
authors.lastname: Amarasena
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, The Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
authors.forename: Sheilajen
authors.initials: S
authors.lastname: Alcantara
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, The Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
authors.forename: Robert
authors.initials: R
authors.lastname: De Rose
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, The Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia; Centre for Biomedical Research, Burnet Institute, Melbourne, Victoria 3004, Australia; Department of Microbiology, Monash University, Clayton, Victoria 3168, Australia.
authors.forename: Gilda
authors.initials: G
authors.lastname: Tachedjian
authors.affiliation: Kirby Institute, University of New South Wales, Sydney, NSW 2052, Australia.
authors.forename: Hamid
authors.initials: H
authors.lastname: Alinejad-Rokny
authors.affiliation: Kirby Institute, University of New South Wales, Sydney, NSW 2052, Australia.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Venturi
authors.affiliation: Kirby Institute, University of New South Wales, Sydney, NSW 2052, Australia.
authors.forename: Miles P
authors.initials: MP
authors.lastname: Davenport
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, The Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
authors.forename: Wendy R
authors.initials: WR
authors.lastname: Winnall
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, The Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia; Melbourne Sexual Health Centre and Department of Infectious Diseases, Alfred Health, Central Clinical School, Monash University, Melbourne, Australia; ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of Melbourne, Parkville, Australia. Electronic address: skent@unimelb.edu.au.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Kent
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D024882:Drug Resistance, Viral / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D008254:Macaca nemestrina; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids / Q000737:chemistry / Q000276:immunology; D012194:RNA-Directed DNA Polymerase / Q000235:genetics* / Q000276:immunology; D016097:Simian Acquired Immunodeficiency Syndrome / Q000276:immunology / Q000821:virology*; D015302:Simian Immunodeficiency Virus / Q000235:genetics* / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology / Q000821:virology; D019562:Viral Load; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology; D014779:Virus Replication / Q000235:genetics*
chemical_list: D014764:Viral Proteins; D012194:RNA-Directed DNA Polymerase
keywords: Drug resistance; Fidelity; Fitness; Human immunodeficiency virus; Nucleotide reverse transcriptase inhibitor; Replication; Reverse transcriptase; Simian immunodeficiency virus