High fidelity simian immunodeficiency virus reverse transcriptase mutants have impaired replication in vitro and in vivo

Sarah B. Lloyd, Marit Lichtfuss, Thakshila H. Amarasena, Sheilajen Alcantara, Robert De Rose, Gilda Tachedjian, Hamid Alinejad-Rokny, Vanessa Venturi, Miles P. Davenport, Wendy R. Winnall, Stephen J. Kent

Research output: Contribution to journalArticleResearchpeer-review

6 Citations (Scopus)

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.
Original languageEnglish
Number of pages10
JournalVirology
Volume492
DOIs
Publication statusPublished - May 2016

Keywords

  • human immunodeficiency virus
  • simian immunodeficiency virus
  • reverse transcriptase
  • fidelity
  • fitness
  • replication
  • nucleotide reverse transcriptase inhibitor
  • drug resistance

Cite this

Lloyd, Sarah B. ; Lichtfuss, Marit ; Amarasena, Thakshila H. ; Alcantara, Sheilajen ; De Rose, Robert ; Tachedjian, Gilda ; Alinejad-Rokny, Hamid ; Venturi, Vanessa ; Davenport, Miles P. ; Winnall, Wendy R. ; Kent, Stephen J. / High fidelity simian immunodeficiency virus reverse transcriptase mutants have impaired replication in vitro and in vivo. In: Virology. 2016 ; Vol. 492.
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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.",
keywords = "human immunodeficiency virus, simian immunodeficiency virus, reverse transcriptase, fidelity, fitness, replication, nucleotide reverse transcriptase inhibitor, drug resistance",
author = "Lloyd, {Sarah B.} and Marit Lichtfuss and Amarasena, {Thakshila H.} and Sheilajen Alcantara and {De Rose}, Robert and Gilda Tachedjian and Hamid Alinejad-Rokny and Vanessa Venturi and Davenport, {Miles P.} and Winnall, {Wendy R.} and Kent, {Stephen J.}",
year = "2016",
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language = "English",
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High fidelity simian immunodeficiency virus reverse transcriptase mutants have impaired replication in vitro and in vivo. / Lloyd, Sarah B.; Lichtfuss, Marit; Amarasena, Thakshila H.; Alcantara, Sheilajen; De Rose, Robert; Tachedjian, Gilda; Alinejad-Rokny, Hamid; Venturi, Vanessa; Davenport, Miles P.; Winnall, Wendy R.; Kent, Stephen J.

In: Virology, Vol. 492, 05.2016.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - High fidelity simian immunodeficiency virus reverse transcriptase mutants have impaired replication in vitro and in vivo

AU - Lloyd, Sarah B.

AU - Lichtfuss, Marit

AU - Amarasena, Thakshila H.

AU - Alcantara, Sheilajen

AU - De Rose, Robert

AU - Tachedjian, Gilda

AU - Alinejad-Rokny, Hamid

AU - Venturi, Vanessa

AU - Davenport, Miles P.

AU - Winnall, Wendy R.

AU - Kent, Stephen J.

PY - 2016/5

Y1 - 2016/5

N2 - 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.

AB - 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.

KW - human immunodeficiency virus

KW - simian immunodeficiency virus

KW - reverse transcriptase

KW - fidelity

KW - fitness

KW - replication

KW - nucleotide reverse transcriptase inhibitor

KW - drug resistance

UR - http://www.ncbi.nlm.nih.gov/pubmed/26896929

U2 - 10.1016/j.virol.2016.02.008

DO - 10.1016/j.virol.2016.02.008

M3 - Article

VL - 492

JO - Virology

JF - Virology

SN - 0042-6822

ER -