A peptide from the GAP-binding domain of the ras-p21 protein as well as azatyrosine block ras-induced maturation of Xenopus oocytes.
The ras-oncogene-encoded p21 protein is known to produce malignant transformation of NIH 3T3 cells as well as maturation of Xenopus oocytes when microinjected into these cells. p21 protein is known to bind a GTPase activating protein (GAP) intracellularly; residues 32-45 have been implicated in interacting with GAP. We demonstrate here that a peptide corresponding to residues 35-47 of p21 as well as the antibiotic azatyrosine inhibit the ras-induced maturation of Xenopus oocytes in a dose-related manner upon microinjection. We have previously shown that this p21 peptide and azatyrosine could inhibit the effects of p21 protein on cell transformation and pinocytosis in NIH 3T3 cells. In the present study, in which we have extended these results to the oocyte system, we also demonstrate that both partially inhibit insulin-induced oocyte maturation, a process which is thought to involve activation of endogenous p21 protein; on the other hand, both agents fail to inhibit oocyte maturation induced by progesterone, which is known not to act through p21 protein activation. Control studies with other peptides and tyrosine analogues support the selective nature of these events. These results suggest that both the p21-related peptide and azatyrosine have potent anti-ras effects intracellularly.
Chung
D L
DL
Department of Chemistry, New York University, NY 10003.
Brandt-Rauf
P
P
Murphy
R B
RB
Nishimura
S
S
Yamaizumi
Z
Z
Weinstein
I B
IB
Pincus
M R
MR
D000409:Alanine / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002454:Cell Differentiation / Q000187:drug effects; D003001:Cloning, Molecular; D005260:Female; D020690:GTPase-Activating Proteins; D005813:Genes, Synthetic; D011905:Genes, ras; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D008969:Molecular Sequence Data; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D010455:Peptides / Q000138:chemical synthesis / Q000494:pharmacology*; D011506:Proteins / Q000378:metabolism*; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics / Q000502:physiology*; D013997:Time Factors; D014982:Xenopus laevis; D020703:ras GTPase-Activating Proteins
D020690:GTPase-Activating Proteins; D007328:Insulin; D010455:Peptides; D011506:Proteins; D020703:ras GTPase-Activating Proteins; C011870:beta-(5-hydroxy-2-pyridyl)alanine; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras); D000409:Alanine