Inhibition of HIV replication by dominant negative mutants of Sam68, a functional homolog of HIV-1 Rev

Nat Med. 1999 Jun;5(6):635-42. doi: 10.1038/9479.

Abstract

The HIV-1 Rev protein facilitates the nuclear export of mRNA containing the Rev response element (RRE) through binding to the export receptor CRM-1. Here we show that a cellular nuclear protein, Sam68 (Src-associated protein in mitosis), specifically interacts with RRE and can partially substitute for as well as synergize with Rev in RRE-mediated gene expression and virus replication. Differential sensitivity to leptomycin B, an inhibitor of CRM-1, indicates that the export pathways mediated by Rev and Sam68 are distinct. C-terminally deleted mutants of Sam68 inhibited the transactivation of RRE-mediated expression by both wild-type Sam68 and Rev. They were retained in the cytoplasm and impeded the nuclear localization of Rev in co-expressed cells. These mutants also inhibited wild-type HIV-1 replication to the same extent as the RevM10 mutant, and may be useful as anti-viral agents in the treatment of AIDS.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Antibodies / metabolism
  • Carrier Proteins / antagonists & inhibitors
  • Carrier Proteins / metabolism
  • Cells, Cultured / virology
  • Chloramphenicol O-Acetyltransferase / genetics
  • Cytoplasm / metabolism
  • DNA-Binding Proteins
  • Enzyme Inhibitors / pharmacology
  • Exportin 1 Protein
  • Fatty Acids, Unsaturated / pharmacology
  • Gene Expression Regulation, Viral
  • Gene Products, rev*
  • Genes, Dominant
  • Genes, Reporter
  • HIV-1 / physiology*
  • HeLa Cells / virology
  • Humans
  • Karyopherins*
  • Kinetin
  • Mutation
  • Purines / pharmacology
  • RNA-Binding Proteins / antagonists & inhibitors
  • RNA-Binding Proteins / genetics*
  • RNA-Binding Proteins / metabolism*
  • Receptors, Cytoplasmic and Nuclear*
  • Response Elements
  • Virus Replication / genetics*
  • rev Gene Products, Human Immunodeficiency Virus

Substances

  • Adaptor Proteins, Signal Transducing
  • Antibodies
  • Carrier Proteins
  • DNA-Binding Proteins
  • Enzyme Inhibitors
  • Fatty Acids, Unsaturated
  • Gene Products, rev
  • KHDRBS1 protein, human
  • Karyopherins
  • Purines
  • RNA-Binding Proteins
  • Receptors, Cytoplasmic and Nuclear
  • rev Gene Products, Human Immunodeficiency Virus
  • olomoucine
  • Chloramphenicol O-Acetyltransferase
  • Kinetin
  • leptomycin B