Genetic variation of the SIVagm transmembrane glycoprotein in naturally and experimentally infected primates.
OBJECTIVE
An in-frame stop codon prematurely truncating the transmembrane glycoprotein (TMP) is a common feature of many simian immunodeficiency virus, African green monkey strain (SIVagm) molecular clones. The purpose of this study was to investigate the native form of the SIVagm TMP in a naturally infected African green monkey (AGM) and to study the fate of the stop codon following the passage of SIVagm in primates.


METHODS
Polymerase chain reaction was used to clone the entire intracellular portion of the TMP from: (1) peripheral blood mononuclear cells (PBMC) of the naturally infected AGM 155; (2) an isolate of SIVagm155 in rhesus PBMC and (3) PBMC from pig-tailed macaques and AGM experimentally infected with an SIVagm molecular clone encoding a truncated TMP.


RESULTS
PBMC of the naturally infected AGM contained a 'swarm' of related virus genotypes that encoded a full-length TMP, whereas tissue-culture passage in rhesus PBMC resulted in a prematurely truncated form of the TMP. This premature stop codon persisted in PBMC of monkeys experimentally infected with an SIVagm molecular clone. Both macaques and AGM of same subspecies as AGM 155 (Cercopithecus pygerythrus) and other subspecies (C. aethiops and C. sabaeus) became infected with SIVagm155. Genetic drift of this region of env, as assessed by calculation of the nucleotide substitution/site/year rate, was similar to that of other retroviruses.


CONCLUSIONS
The native form of the SIVagm TMP is a full-length gp40, similar to the SIV macaque (SIVmac) strain and HIV-1. However, passage of SIVagm in tissue culture can result in point mutations that introduce a premature stop codon. This stop codon persists during subsequent in vivo passage of SIVagm in primates. This contrasts with similar studies in macaques infected with SIVmac, in which reversion of the TMP stop codon was observed.
Department of Microbiology, Georgetown University, Rockville, Maryland.
A
A
Fomsgaard
P R
PR
Johnson
W T
WT
London
V M
VM
Hirsch
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D003062:Codon; D017931:DNA Primers; D015686:Gene Products, env / Q000235:genetics*; D014644:Genetic Variation; D006801:Humans; D008253:Macaca mulatta; D008254:Macaca nemestrina; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D015302:Simian Immunodeficiency Virus / Q000145:classification / Q000235:genetics*; D013728:Terminator Regions, Genetic; D014760:Viral Fusion Proteins
D003062:Codon; D017931:DNA Primers; D015686:Gene Products, env; D015682:Retroviridae Proteins, Oncogenic; D014760:Viral Fusion Proteins; C059580:transmembrane protein, Simian immunodeficiency virus