Regulation of specific DNA binding by p53: evidence for a role for O-glycosylation and charged residues at the carboxy-terminus.

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State: Public
Version: Final published version
License: All rights reserved
Serval ID
serval:BIB_2683E03B3010
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Regulation of specific DNA binding by p53: evidence for a role for O-glycosylation and charged residues at the carboxy-terminus.
Journal
Oncogene
Author(s)
Shaw P., Freeman J., Bovey R., Iggo R.
ISSN
0950-9232 (Print)
ISSN-L
0950-9232
Publication state
Published
Issued date
15/02/1996
Peer-reviewed
Oui
Volume
12
Number
4
Pages
921-930
Language
english
Notes
Publication types: Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
The carboxy-terminus of p53 contains a basic region which represses DNA binding, and this repression can be relieved by PAb421, an antibody against the basic region. The EB-1 human cell line contains wild type p53 protein which fails to express the PAb421 epitope and is highly active both in biological assays and in DNA binding assays. We show by wheat germ agglutinin chromatography and galactosyl-transferase labelling that this p53 is O-glycosylated, and that at least one of the sugar residues masks the PAb421 epitope, as demonstrated by recovery of reactivity with PAb421 after digestion of Western blots of EB-1 cell extract with hexosaminidase. A minor population of p53 molecules in EB-1 cells lacks the modification, and there is a correlation between the ability to bind DNA with high affinity and masking of the PAb421 epitope. We also show that strongly positively charged peptides, including short peptides from the basic region of p53, can derepress DNA binding, probably by disruption of an intramolecular interaction involving the basic region. We propose that any intervention which prevents this intramolecular interaction, including addition of bulky residues such as sugar groups, can activate DNA binding by p53.
Keywords
Amino Acid Sequence, Base Sequence, Blotting, Western, Cell Line, Cloning, Molecular, DNA/metabolism, DNA Primers, DNA-Binding Proteins/isolation & purification, DNA-Binding Proteins/metabolism, Epitopes/analysis, Epitopes/chemistry, Glycosylation, Humans, Molecular Sequence Data, Oligonucleotide Probes, Polymerase Chain Reaction, Recombinant Proteins/biosynthesis, Recombinant Proteins/isolation & purification, Recombinant Proteins/metabolism, Saccharomyces cerevisiae, Sequence Homology, Amino Acid, Tumor Suppressor Protein p53/biosynthesis, Tumor Suppressor Protein p53/isolation & purification, Tumor Suppressor Protein p53/metabolism
Pubmed
Web of science
Create date
29/01/2008 18:36
Last modification date
16/07/2020 8:42
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