Document 0169
 DOCN  M94A0169
 TI    Construction and in vitro properties of SIVmac mutants with deletions in
       nonessential genes.
 DT    9412
 AU    Gibbs JS; Regier DA; Desrosiers RC; Harvard Medical School, New England
       Regional Primate Research; Center, Southborough, Massachusetts
       01772-9102.
 SO    AIDS Res Hum Retroviruses. 1994 Apr;10(4):333-42. Unique Identifier :
       AIDSLINE MED/94347457
 AB    The so-called nonessential genes of primate lentiviruses can be deleted
       without abrogating the ability of virus to replicate under at least some
       cell culture conditions. In SIVmac, these genes are vif, vpx, vpr, and
       nef. Sequences in the upstream region of U3 in the LTR have also been
       shown to be dispensable for replication in cell culture. We report here
       the construction and characterization of a panel of 40 single and
       combination deletion mutants derived from the pathogenic molecular clone
       SIVmac239. Deletion of the vpr gene caused little or no change in the
       growth properties of SIVmac239 in CEMx174 cells, in rhesus monkey
       peripheral blood mononuclear cells (PBMCs), or in rhesus monkey alveolar
       macrophages. Deletion of the vpx gene resulted in a greatly reduced rate
       of replication of the virus in the primary PBMC and macrophage cultures,
       but no significant reduction in replication of the virus in CEMx174
       cells. Deletion of the vpx gene appeared to have a greater effect on
       virus replication in macrophages than in PBMCs. Deletion of the vif gene
       caused a dramatic reduction in replication in all cell types tested.
       However, even delta 5, which contains deletions in all five targeted
       regions (vif, vpx, vpr, nef, and U3), can still replicate in CEMx174
       cells albeit with greatly delayed kinetics. Deletion of nef, alone or in
       combination with deletions in U3 and vpr, had no observable effect on
       replication of the virus in any of the cells tested. Because the disease
       induced by the parental SIVmac239 clone in rhesus monkeys has been well
       characterized and is remarkably similar to AIDS in humans, this
       collection of mutants will be useful for relating in vitro properties
       and gene function with in vivo pathogenic potential.
 DE    Amino Acid Sequence  Animal  Base Sequence  Cell Line  DNA
       Primers/GENETICS  DNA, Viral/GENETICS  *Gene Deletion  Genes, nef
       Genes, vif  Genes, vpr  *Genes, Viral  In Vitro  Leukocytes,
       Mononuclear/MICROBIOLOGY  Macaca mulatta  Macrophages,
       Alveolar/MICROBIOLOGY  Molecular Sequence Data  Mutagenesis,
       Site-Directed  Support, U.S. Gov't, P.H.S.
       SIV/*GENETICS/PHYSIOLOGY/PATHOGENICITY  Viral Proteins/GENETICS  Virus
       Replication/GENETICS  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).