A carboxy-terminally truncated form of the Vpr protein of human immunodeficiency virus type 1 retards cell proliferation independently of G(2) arrest of the cell cycle.
Vpr, one of the accessory gene products of HIV-1, is a 96-residue protein with several functions. It is involved in import of the HIV-1 preintegration complex into the nucleus of nondividing cells, in cellular differentiation, inducing cell cycle arrest at the G(2)/M phase, in immune suppression, and in enhancement of replication of the virus. We found recently that Vpr interferes with the proliferation of mouse NIH3T3 fibroblasts but fails to arrest these cells in the G(2) phase. Thus, it seems possible that Vpr might retard cell proliferation via a novel pathway that is distinct from G(2) arrest. To elucidate the mechanism by which Vpr induces the retardation of cell growth, we developed a panel of expression vectors that encoded Vpr molecules with deletions of specific putative domains, namely, the first alpha-helical domain, the second alpha-helical domain, a leucine zipper-like domain, and an arginine-rich carboxy-terminal domain. These vectors were introduced into HeLa cells since expression of Vpr can induce G(2) arrest in such cells. A carboxy-terminally truncated form of Vpr, C81, which failed to induce G(2) arrest, led to the G(1) arrest and retained the ability to prevent cell proliferation. All the other mutant proteins had completely lost the capacity to induce G(2) arrest and to suppress growth. Substitutions of Ile/Leu for Pro at positions 60, 67, 74, and 81 within the leucine zipper-like domain of Vpr or of C81 revealed that Ile60, Leu67, and Ile74 play an important role in the C81-induced suppression of growth, while Ile74 and Ile81 were found to be indispensable for Vpr-induced G(2) arrest. Collectively, our results strongly suggest that Vpr can retard cell proliferation independently of G(2) arrest of the cell cycle.
Nishizawa
M
M
Tsukuba Life Science Center, The Institute of Physical and Chemical Research (RIKEN), 3-1-1 Koyadai, Tsukuba, Ibaraki, 305-0074, Japan.
Myojin
T
T
Nishino
Y
Y
Nakai
Y
Y
Kamata
M
M
Aida
Y
Y
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D002453:Cell Cycle; D002455:Cell Division; D004247:DNA / Q000032:analysis / Q000096:biosynthesis; D016195:G2 Phase; D016336:Gene Products, vpr / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D016350:Leucine Zippers; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D015398:Signal Transduction; D014162:Transfection; D054325:vpr Gene Products, Human Immunodeficiency Virus
D016336:Gene Products, vpr; D011993:Recombinant Fusion Proteins; D054325:vpr Gene Products, Human Immunodeficiency Virus; D004247:DNA