title: Slower uncoating is associated with impaired replicative capability of simian-tropic HIV-1.
abstract: Human immunodeficiency virus type 1 (HIV-1) productively infects only humans and chimpanzees, but not Old World monkeys, such as rhesus and cynomolgus (CM) monkeys. To establish a monkey model of HIV-1/AIDS, several HIV-1 derivatives have been constructed. We previously generated a simian-tropic HIV-1 that replicates efficiently in CM cells. This virus encodes a capsid protein (CA) with SIVmac239-derived loops between α-helices 4 and 5 (L4/5) and between α-helices 6 and 7 (L6/7), along with the entire vif from SIVmac239 (NL-4/5S6/7SvifS). These SIVmac239-derived sequences were expected to protect the virus from HIV-1 restriction factors in monkey cells. However, the replicative capability of NL-4/5S6/7SvifS in human cells was severely impaired. By long-term cultivation of human CEM-SS cells infected with NL-4/5S6/7SvifS, we succeeded in partially rescuing the impaired replicative capability of the virus in human cells. This adapted virus encoded a G-to-E substitution at the 116(th) position of the CA (NL-4/5SG116E6/7SvifS). In the work described here, we explored the mechanism by which the replicative capability of NL-4/5S6/7SvifS was impaired in human cells. Quantitative analysis (by real-time PCR) of viral DNA synthesis from infected cells revealed that NL-4/5S6/7SvifS had a major defect in nuclear entry. Mutations in CA are known to affect viral core stability and result in deleterious effects in HIV-1 infection; therefore, we measured the kinetics of uncoating of these viruses. The uncoating of NL-4/5S6/7SvifS was significantly slower than that of wild type HIV-1 (WT), whereas the uncoating of NL-4/5SG116E6/7SvifS was similar to that of WT. Our results suggested that the lower replicative capability of NL-4/5S6/7SvifS in human cells was, at least in part, due to the slower uncoating of this virus.
authors: Department of Viral Infections, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan.
authors: Ken
authors: K
authors: Kono
authors: Eri
authors: E
authors: Takeda
authors: Hiromi
authors: H
authors: Tsutsui
authors: Ayumu
authors: A
authors: Kuroishi
authors: Amy E
authors: AE
authors: Hulme
authors: Thomas J
authors: TJ
authors: Hope
authors: Emi E
authors: EE
authors: Nakayama
authors: Tatsuo
authors: T
authors: Shioda
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics; D002460:Cell Line; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D054303:HIV Reverse Transcriptase / Q000378:metabolism; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D009154:Mutation; D016097:Simian Acquired Immunodeficiency Syndrome / Q000821:virology; D015302:Simian Immunodeficiency Virus / Q000502:physiology*; D013997:Time Factors; D014779:Virus Replication; D057646:Virus Uncoating
chemical_list: D036022:Capsid Proteins; D054303:HIV Reverse Transcriptase
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