Wild-type alternatively spliced p53

Binding to DNA and interaction with the major p53 protein in vitro and in cells

Yu Wu, Yuangang Liu, Laura Lee, Zoe Miner, Molly Kulesz-Martin

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

A p53 variant protein (p53as) generated from alternatively spliced p53 RNA is expressed in normal and malignant mouse cells and tissues, and p53as antigen activity is preferentially associated with the G2 phase of the cell cycle, suggesting that p53as and p53 protein may have distinct properties. Using p53as and p53 proteins translated in vitro, we now provide evidence that p53as protein has efficient sequence-specific DNA-binding ability. DNA binding by p53 protein is inefficient in comparison and requires activation. Furthermore, p53as and p53 proteins formed hetero-oligomers when co-translated in vitro, resulting in inactivation of p53as DNA-binding activity. Gel filtration indicated that p53as translated in vitro, like p53, formed tetramers. In support of a functional role of p53as in cells, p53as/p53 hetero-oligomers were co-immunoprecipitated from mouse cells, and both protein forms were detectable in nuclear extracts by electrophoretic mobility shift assays. These results suggest that the biochemical functions of p53 are mediated by interaction between two endogenous protein products of the wild-type p53 gene.

Original languageEnglish (US)
Pages (from-to)4823-4830
Number of pages8
JournalEMBO Journal
Volume13
Issue number20
StatePublished - 1994
Externally publishedYes

Fingerprint

DNA
Proteins
Oligomers
G2 Phase
p53 Genes
Electrophoretic mobility
DNA-Binding Proteins
Electrophoretic Mobility Shift Assay
In Vitro Techniques
Gel Chromatography
Cell Cycle
Assays
RNA
Genes
Gels
Chemical activation
Antigens
Cells
Tissue

Keywords

  • Alternative splicing
  • DNA binding
  • Wild-type p53 protein

ASJC Scopus subject areas

  • Cell Biology
  • Genetics

Cite this

Wild-type alternatively spliced p53 : Binding to DNA and interaction with the major p53 protein in vitro and in cells. / Wu, Yu; Liu, Yuangang; Lee, Laura; Miner, Zoe; Kulesz-Martin, Molly.

In: EMBO Journal, Vol. 13, No. 20, 1994, p. 4823-4830.

Research output: Contribution to journalArticle

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