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PMID: 9094635 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Regulation of p53-dependent apoptosis, transcriptional repression, and cell transformation by phosphorylation of the 55-kilodalton E1B protein of human adenovirus type 5.

Journal of virology ·Vol. 71 ·No. 5 ·1997-05-00 ·Pages 3620-7

Teodoro JG, Branton PE

Abstract

The adenovirus type 5 55-kDa E1B protein (E1B-55kDa) cooperates with E1A gene products to induce cell transformation. E1A proteins stimulate DNA synthesis and cell proliferation; however, they also cause rapid cell death by p53-dependent and p53-independent apoptosis. It is believed that the role of the E1B-55kDa protein in transformation is to protect against p53-dependent apoptosis by binding to and inactivating p53. It has been shown previously that the 55-kDa polypeptide abrogates p53-mediated transactivation and that mutants defective in p53 binding are unable to cooperate with E1A in transformation. We have previously mapped phosphorylation sites near the carboxy terminus of the E1B-55kDa protein at Ser-490 and Ser-491, which lie within casein kinase II consensus sequences. Conversion of these sites to alanine residues greatly reduced transforming activity, and although the mutant 55-kDa protein was found to interact with p53 at normal levels, it was somewhat defective for suppression of p53 transactivation activity. We now report that a nearby residue, Thr-495, also appears to be phosphorylated. We demonstrate directly that the wild-type 55-kDa protein is able to block E1A-induced p53-dependent apoptosis, whereas cells infected by mutant pm490/1/5A, which contains alanine residues at all three phosphorylation sites, exhibited extensive DNA fragmentation and classic apoptotic cell death. The E1B-55kDa product has been shown to exhibit intrinsic transcriptional repression activity when localized to promoters, such as by fusion with the GAL4 DNA-binding domain, even in the absence of p53. Such repression activity was totally absent with mutant pm490/1/5A. These data suggested that inhibition of p53-dependent apoptosis may depend on the transcriptional repression function of the 55-kDa protein, which appears to be regulated be phosphorylation at the carboxy terminus.

MeSH Terms
Adenovirus E1B Proteins/physiology Adenoviruses, Human/chemistry Animals Apoptosis Cell Transformation, Neoplastic Humans KB Cells Mice Molecular Weight Phosphorylation Transcription, Genetic Tumor Suppressor Protein p53/physiology
Chemicals
Adenovirus E1B Proteins Tumor Suppressor Protein p53
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Teodoro J G
Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
Branton P E
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.71.5.3620-3627.1997
Published
1997-05-00
Pages
3620-7
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC191510
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.71.5.3620-3627.1997 PMC Full Text View journal details
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