Mutational analysis of p50E4F suggests that DNA binding activity is mediated through an alternative structure in a zinc finger domain that is regulated by phosphorylation

Robert Rooney, Kristen Rothammer, Elma R. Fernandes

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

p50E4F is a cellular transcription factor whose DNA binding activity is stimulated in a phosphorylation-dependent manner by products of the adenovirus E1A oncogene. Although p50E4F does not contain a bZIP DNA binding motif, it binds a tandemly repeated palindromic sequence in the adenovirus E4 promoter that is recognized by a large number of bZIP proteins, but with much greater stability. Analysis of deletions in the p50E4F sequence identified the regions that are responsible for its unique DNA binding properties. Sequence-specific DNA binding and factor dimerization were localized to a C-terminal region containing two C2H2 and one CCHC zinc finger motifs; the phosphorylation site critical for DNA binding activity was also localized to this domain. The high stability of p50E4F binding also required residues within the first 83 amino acids of the N-terminus. Analysis of single and double amino acid substitutions in the C-terminal zinc finger domain demonstrated that while the second C2H2 zinc finger was required for DNA binding activity, the putative structures of the first C2H2 and the CCHC zinc fingers were not. Instead, residues from these other zinc finger motifs appeared to participate in an alternative structure that mediates DNA binding activity and is regulated by phosphorylation.

Original languageEnglish (US)
Pages (from-to)1681-1688
Number of pages8
JournalNucleic acids research
Volume26
Issue number7
DOIs
StatePublished - Apr 1 1998

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Zinc Fingers
Phosphorylation
DNA
Adenoviridae
Basic-Leucine Zipper Transcription Factors
Nucleotide Motifs
Dimerization
Amino Acid Substitution
Oncogenes
Transcription Factors
Amino Acids
CYS2-HIS2 Zinc Fingers

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Mutational analysis of p50E4F suggests that DNA binding activity is mediated through an alternative structure in a zinc finger domain that is regulated by phosphorylation. / Rooney, Robert; Rothammer, Kristen; Fernandes, Elma R.

In: Nucleic acids research, Vol. 26, No. 7, 01.04.1998, p. 1681-1688.

Research output: Contribution to journalArticle

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