Biophysical characterizations of human mitochondrial transcription factor A and its binding to tumor suppressor p53

Tuck Seng Wong, Sridharan Rajagopalan, Stefan M. Freund, Trevor J. Rutherford, Antonina Andreeva, Fiona M. Townsley, Miriana Petrovich, Alan R. Fersht*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

87 Citations (Scopus)

Abstract

Human mitochondrial transcription factor A (TFAM) is a multi-functional protein, involved in different aspects of maintaining mitochondrial genome integrity. In this report, we characterized TFAM and its interaction with tumor suppressor p53 using various biophysical methods. DNA-free TFAM is a thermally unstable protein that is in equilibrium between monomers and dimers. Self-association of TFAM is modulated by its basic C-terminal tail. The DNA-binding ability of TFAM is mainly contributed by its first HMG-box, while the second HMG-box has low-DNA-binding capability. We also obtained backbone resonance assignments from the NMR spectra of both HMG-boxes of TFAM. TFAM binds primarily to the N-terminal transactivation domain of p53, with a Kd of 1.95 ± 0.19 μM. The C-terminal regulatory domain of p53 provides a secondary binding site for TFAM. The TFAM-p53-binding interface involves both TAD1 and TAD2 sub-domains of p53. Helices α1 and α2 of the HMG-box constitute the main p53-binding region. Since both TFAM and p53 binds preferentially to distorted DNA, the TFAM-p53 interaction is implicated in DNA damage and repair. In addition, the DNA-binding mechanism of TFAM and biological relevance of the TFAM-p53 interaction are discussed.

Original languageEnglish
Article numbergkp750
Pages (from-to)6765-6783
Number of pages19
JournalNucleic Acids Research
Volume37
Issue number20
DOIs
Publication statusPublished - 15 Sept 2009

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