Mutational analysis of HIV-1 Nef: identification of two mutants that are temperature-sensitive for CD4 downregulation

Abstract

The Nef protein of HIV-1 is capable of performing at least two functions: reduction of cell-surface expression of CD4, the viral receptor, and enhancement of HIV-1 infectivity. Here we report the results of assays of CD4 downregulation by nef point mutants. Areas of high charge density and highly conserved sequence motifs were targeted. In general, mutations in the carboxyl-terminal half of the protein had more deleterious effects than did those in the amino-terminal half. A single mutant nef allele was identified which encoded a stable protein but was completely inactive. Two mutant Nef proteins exhibited temperature-sensitive phenotypes in assays of CD4 downregulation.