Protein-protein interactions in high molecular weight forms of the transformation-related phosphoprotein p53.
The transformation-related cellular phosphoprotein p53 interacts with a variety of viral and cellular proteins and with itself to form high molecular weight complexes. The formation of high molecular weight complexes correlates with the transformed morphology of the cells whereas in non-transformed cells low molecular weight forms are predominant. Thus, aggregation seems to be involved in the regulation of biological functions of p53. Analyzing wild-type and mutant p53 in the same cellular environment i.e. after an in vitro transcription/translation reaction in rabbit reticulocytes we found high molecular weight forms for wild-type and mutant p53. The sedimentation profile resembled the profile obtained for mutant p53 from transformed cells. As shown by dilution experiments, aggregation of p53 was not due to high p53 protein concentrations. Although p53 is known to bind RNA, treatment with RNAse did not change the aggregation state of p53 suggesting that RNA may not contribute to the quaternary structure of p53. High molecular weight aggregates of p53 were resistant to treatment with 1 M NaCl and also stable in weak acidic conditions. Alkaline pH as well as treatment with 3.5 M NaCl led to a disaggregation of high molecular weight complexes of p53. This treatment resulted in low molecular weight forms consisting probably of dimers to tetramers whereas monomers of p53 are hardly detectable. A nearly complete disaggregation was obtained only with the ionic denaturing detergent sodium dodecyl sulfate. Therefore, one has to assume different types of protein-protein interactions leading to the various quaternary structures of p53.
Kraiss
S
S
Department of Biochemistry, University of Ulm, Germany.
Lorenz
A
A
Montenarh
M
M
D000818:Animals; D002461:Cell Line, Transformed; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D016158:Genes, p53; D011905:Genes, ras; D046911:Macromolecular Substances; D008970:Molecular Weight; D014176:Protein Biosynthesis; D011487:Protein Conformation; D051381:Rats; D014158:Transcription, Genetic; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification
D046911:Macromolecular Substances; D016159:Tumor Suppressor Protein p53