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PMID: 8013454 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A temperature-sensitive mutant of human p53.

The EMBO journal ·Vol. 13 ·No. 11 ·1994-06-01 ·Pages 2535-44

Zhang W, Guo XY, Hu GY, Liu WB, Shay JW, Deisseroth AB

Abstract

We discovered that the human 'hot-spot' p53 mutant 143Ala is temperature sensitive for the binding of two DNA elements and for mediating the transcription of a downstream luciferase reporter gene. At 32.5 degrees C, 143Ala possesses strong DNA binding ability. In addition, its transcriptional activities are stronger than those of wild-type p53. At 37.5 degrees C, however, both the mutant's DNA binding and transcriptional activation functions are greatly reduced or abolished. The activity differences correlate with the presence of two conformational states of p53, which are recognized by monoclonal antibodies: at 32.5 degrees C, 143Ala is recognized by PAb1620, which is specific for the wild-type conformation, whereas at 37.5 degrees C, 143Ala is almost undetectable by PAb1620-mediated immunoprecipitation. Although 143Ala's binding to p53 DNA recognition elements and its activation of reporter gene transcription at 32.5 degrees C is markedly higher than that of the wild-type p53, 143Ala inhibited proliferation less robustly than wild-type p53 and it did not increase inhibition of ras-induced focus formation. These results indicate a partial correlation between the events involved in transcriptional activation and events involved in the proliferation suppression of p53. 143Ala therefore will be of use in dissecting the relationship between the structure of p53 and its different cellular functions.

Related Genes
p53
MeSH Terms
Animals Antibodies, Monoclonal Cell Division Cell Transformation, Neoplastic DNA/metabolism Genes, Reporter Genes, p53/genetics Genes, ras Humans Mice Mutation/genetics Protein Binding Protein Conformation Rats Regulatory Sequences, Nucleic Acid/genetics Structure-Activity Relationship Temperature Transcription, Genetic Transcriptional Activation/genetics Tumor Cells, Cultured Tumor Suppressor Protein p53/chemistry,genetics,metabolism,physiology
Chemicals
Antibodies, Monoclonal Tumor Suppressor Protein p53 DNA
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Zhang W
Department of Hematology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
Guo X Y
Hu G Y
Liu W B
Shay J W
Deisseroth A B
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1994-06-01
Pages
2535-44
Language
English
Region
England
NLM ID
8208664
PMCID
PMC395127
Subset
IM
Grants
NIA NIH HHS · AG07992 · United States
NCI NIH HHS · CA50195 · United States
NCI NIH HHS · P01 CA55164 · United States
External Links
PubMed source PMC Full Text View journal details
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