RNA-p53 interactions in vitro

Research output: Contribution to journalArticle

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Abstract

The tumor suppressor protein p53 is mutated in over half of human cancers. Despite 25 years of study, the complex regulation of this protein remains unclear. After serendipitously detecting RNA binding by p53 in the yeast three-hybrid system (Y3H), we are exploring the specificity and function of this interaction. Electrophoretic mobility shift assays show that full-length p53 binds equally to RNAs that are strongly distinguished in the Y3H. RNA binding blocks sequence-specific DNA binding by p53. The C-terminus of p53 is necessary and sufficient for strong RNA interaction in vitro. Mouse and human C-terminal p53 peptides have different affinities for RNA, and an acetylated human p53 C-terminal peptide does not bind RNA. Circular dichroism spectroscopy of p53 peptides shows that RNA binding does not induce a structural change in the p53 C-terminal peptide, and C-terminal peptides do not detectably affect the structure of RNA. These results demonstrate that p53 binds RNA with little sequence specificity, RNA binding has the potential to regulate DNA binding, and RNA-p53 interactions can be regulated by acetylation of the p53 C-terminus.

Original languageEnglish (US)
Pages (from-to)2480-2487
Number of pages8
JournalBiochemistry
Volume46
Issue number9
DOIs
StatePublished - Mar 6 2007

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RNA
Peptides
In Vitro Techniques
Tumor Suppressor Protein p53
Circular dichroism spectroscopy
Two-Hybrid System Techniques
Acetylation
Electrophoretic mobility
Electrophoretic Mobility Shift Assay
Circular Dichroism
DNA
Hybrid systems
Yeast
Spectrum Analysis
Assays
Neoplasms
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

RNA-p53 interactions in vitro. / Riley, Kasandra J L; Ramirez-Alvarado, Marina; Maher III, L James.

In: Biochemistry, Vol. 46, No. 9, 06.03.2007, p. 2480-2487.

Research output: Contribution to journalArticle

Riley, Kasandra J L ; Ramirez-Alvarado, Marina ; Maher III, L James. / RNA-p53 interactions in vitro. In: Biochemistry. 2007 ; Vol. 46, No. 9. pp. 2480-2487.
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