Important note: Information in this article was accurate in 1991. The state of the art may have changed since the publication date.
Nucleotide sequence of the pathogenic SIV239 clone and analysis of nef function.
Symp Nonhum Primate Models AIDS. 1990 Nov 28-30;8:21 (abstract no. 5). Unique Identifier : AIDSLINE PRIM8/900005 Kestler H; Regier D; Mori K; Kodama T; Daniel M; Desrosiers R; Department of Microbiology and Molecular Genetics, New England; Regional Primate Research Center, Harvard Medical School,; Southborough, MA, USA
Abstract:
The SIVmac239 infectious molecular clone yields virus that induces AIDS in rhesus monkeys in a time frame suitable for laboratory investigation. The proviral genome, including both LTRs, is 10,279 base pairs. Nine open reading frames, previously identified in other HIV-2/SIVmac/SIVsmm isolates, were also present in this clone. The reading frame for the transmembrane protein is fully open and produces a 41 kilodalton polypeptide in macaque PBL cultures. The nef gene is truncated by the presence of a premature stop codon 93 amino acids from the start of the nef reading frame. Site specific mutagenesis was used to create forms of this clone with a fully open nef reading frame (nef-open) and with a 172 bp deletion in nef (nef-deletion). These three forms of cloned SIVmac239 are otherwise isogenic. The three forms of virus replicated with very similar kinetics and extents in CEMX174 cells and in macaque PBL following transfection. Rhesus monkeys became infected following inoculation with all three forms of cloned SIVmac239. We isolated five DNA clones that span nef sequences at 20, 69 and 93 weeks following infection of two rhesus monkeys with SIVmac239/nef-stop. In all five cases, the stop codon in nef had reverted to a coding codon. Thus, open forms of nef are selected during in vivo infection. Virus loads were at least 100 to 1,000 times lower in rhesus monkeys that were infected with SIVmac239/nef-deletion than in monkeys that received nef+ SIVmac239. None of the six animals that received SIVmac239/nef-deletion have at this time developed AIDS while 5 of 12 animals that received SIVmac239 virus with functional nef genes have died with AIDS. These findings indicate that the open form of nef in SIVmac239 is a functional one and can be used to analyze the role of nef in viral infection. Furthermore, our results suggest that nef is critical for the virus life cycle in vivo, and is required for full pathogenic potential.
Keywords: Animal Base Sequence Cells, Cultured Chromosome Deletion Clone Cells Codon *Genes, nef HIV Long Terminal Repeat HIV-2/GENETICS Kinetics Macaca mulatta Mutagenesis, Site-Directed Open Reading Frames Simian Acquired Immunodeficiency Syndrome/GENETICS SIV/GROWTH & DEVELOPMENT/*GENETICS Transfection Virus Replication ABSTRACT 910730
M9170989
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Important note: Information in this article was accurate in 1991. The state of the art may have changed since the publication date.
