A complex promoter element mediates transactivation of the human proliferating cell nuclear antigen promoter by the 243-residue adenovirus E1A oncoprotein

Labrie, C., Morris, G. F., Mathews, M. B. (March 1993) A complex promoter element mediates transactivation of the human proliferating cell nuclear antigen promoter by the 243-residue adenovirus E1A oncoprotein. Mol Cell Biol, 13 (3). pp. 1697-707. ISSN 0270-7306 (Print)

URL: http://www.ncbi.nlm.nih.gov/pubmed/8095093
DOI: 10.1128/MCB.13.3.1697

Abstract

The adenovirus E1A oncoproteins interfere with the normal regulation of cellular proliferation through interactions with cell cycle regulatory proteins. In view of the essential role of proliferating-cell nuclear antigen (PCNA) in DNA replication, we performed a mutational analysis of the minimal human PCNA promoter (nucleotides -87 to +62) to define sequence elements which mediate transactivation by the 243-residue E1A protein (E1A 243R). Linker-scanning and site-directed mutants were examined for basal and E1A-induced expression of chloramphenicol acetyltransferase (CAT) from PCNA promoter-CAT reporter constructs transiently expressed in HeLa cells. The results define the cis-acting element required for induction of PCNA by E1A 243R as a region between -59 and -45 relative to the transcription initiation site. This PCNA E1A-responsive element (PERE), which is protected from DNase I digestion by nuclear extracts from 293 cells, includes the sequence AGCGTGG immediately upstream of the ATF binding site previously shown to be important for activation of PCNA by E1A 243R (G. F. Morris and M. B. Mathews, J. Virol. 65:6397-6406, 1991). Mutation of either the upstream component or the ATF site within the PERE diminishes basal promoter activity and abrogates transactivation by E1A 243R. This novel cis-acting element is also essential for both basal and E1A-induced expression in the context of the full-length PCNA promoter.

Item Type: Paper
Uncontrolled Keywords: Adenovirus E1A Proteins/ metabolism Antigens, Neoplasm/biosynthesis/ genetics Base Sequence Binding Sites Comparative Study Consensus Sequence DNA Mutational Analysis Hela Cells Humans Molecular Sequence Data Mutagenesis Nuclear Proteins/biosynthesis/ genetics/metabolism Proliferating Cell Nuclear Antigen Promoter Regions (Genetics)/ genetics Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Sequence Alignment Trans-Activation (Genetics) Transcription, Genetic Variation (Genetics)
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > transcription
organism description > virus > adenovirus
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > mutations > mutagenesis
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > Proliferating Cell Nuclear Antigen
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA expression > promoter
CSHL Authors:
Communities: CSHL labs
Depositing User: Matt Covey
Date: March 1993
Date Deposited: 13 Apr 2016 15:25
Last Modified: 13 Apr 2016 15:25
PMCID: PMC359482
Related URLs:
URI: http://repository.cshl.edu/id/eprint/32560

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