A Proteolytic Fragment from the Central Region of P53 Has Marked Sequence-Specific DNA-Binding Activity When Generated from Wild-Type but Not from Oncogenic Mutant P53-Protein

Bargonetti, J., Manfredi, J. J., Chen, X. B., Marshak, D. R., Prives, C. (December 1993) A Proteolytic Fragment from the Central Region of P53 Has Marked Sequence-Specific DNA-Binding Activity When Generated from Wild-Type but Not from Oncogenic Mutant P53-Protein. Genes Dev, 7 (12B). pp. 2565-2574. ISSN 0890-9369

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Abstract

p53 is a sequence-specific DNA-binding oligomeric protein that can activate transcription from promoters bearing p53-binding sites. Whereas the activation region of p53 has been identified within the amino terminus, the location of the specific DNA-binding domain has not been reported. Thermolysin treatment of p53 protein generates a stable protease-resistant fragment that binds with marked specificity to p53 DNA-binding sites. Amino-terminal sequencing of the fragment located the thermolysin cleavage site to residue 91. Because the fragment does not contain the cdc2 phosphorylation site at Ser-315, we conclude that the the site-specific DNA-binding domain of p53 spans the central region of the protein. The vast majority of the mutations in oncogenically derived p53 proteins are located within this central portion of the molecule. Such mutant p53 proteins exhibit defective sequence-specific DNA-binding. Although thermolysin digestion of mutant p53 proteins generates proteolytic patterns that differ from wild-type protein, one mutant tested, His-273, generates a resistant' fragment that migrates with a similar electrophoretic mobility to the wild-type protease-resistant fragment. Interestingly, although intact mutant His-273 protein binds to DNA at 20-degrees-C, the thermolysin-resistant mutant fragment does not. In addition, the central protease-resistant, site-specific binding region of wild-type p53 does not demonstrate nonspecific DNA-binding. Thus, although sequences outside of the central region of p53 contribute to both nonspecific DNA-binding and oligomerization, they are not required for sequence-specific DNA-binding.

Item Type:	Paper
Uncontrolled Keywords:	P53 DNA-BINDING DOMAIN SEQUENCE-SPECIFIC DNA BINDING TUMOR SUPPRESSOR PROTEIN CELL-CYCLE CHECKPOINT CASEIN KINASE-II LARGE T-ANTIGEN MONOCLONAL-ANTIBODIES GENE-EXPRESSION IDENTIFICATION MUTATIONS INVITRO DOMAIN
Subjects:	bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > DNA binding protein bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > mutations bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > genes: types > oncogene bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > genes: types > p53
CSHL Authors:	Marshak, Daniel R.
Communities:	CSHL labs
Depositing User:	Matt Covey
Date:	December 1993
Date Deposited:	20 Apr 2016 18:50
Last Modified:	03 Nov 2017 20:29
Related URLs:	Publisher
URI:	https://repository.cshl.edu/id/eprint/32510

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