HIV-1 Nef protein causes death in stressed yeast cells due to determinants near the N-terminus and elsewhere in Nef.
The Human Immunodeficiency Virus type 1 (HIV-1) Nef protein is essential for AIDS pathogenesis. In order to determine more about the effects of Nef on basic cellular functions Nef was produced in yeast under a variety of conditions and in multiple cell types. Production of Nef caused cell death in acutely copper- or heat-stressed diploid cells. The N-terminal melittin-like region of Nef was involved in toxicity since a Trp5-->Ala change within Nef change caused increased toxicity. However, another determinant was also involved in toxicity since production of Nef20-206 was also still toxic. In each of these Nef-producing cells there was coincident membrane permeabilisation. These results suggest the possibility of a novel yeast bioassay for Nef inhibitors and that cells producing high levels of Nef may be selectively killed by stress.
Macreadie
I
I
Biomolecular Research Institute, Parkville, Victoria, Australia.
Sankovich
S
S
Failla
P
P
Lowe
M
M
Curtain
C
C
Castelli
L
L
Azad
A
A
D000595:Amino Acid Sequence; D002463:Cell Membrane Permeability; D019327:Copper Sulfate / Q000494:pharmacology; D003470:Culture Media; D015693:Gene Products, nef / Q000737:chemistry / Q000502:physiology*; D015754:Genes, nef; D015497:HIV-1; D006358:Hot Temperature; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000254:growth & development* / Q000502:physiology; D014170:Transformation, Genetic; D054311:nef Gene Products, Human Immunodeficiency Virus
D003470:Culture Media; D015693:Gene Products, nef; D054311:nef Gene Products, Human Immunodeficiency Virus; D019327:Copper Sulfate