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Journal of Biochemistry Advance Access published online on December 13, 2006
Journal of Biochemistry, doi:10.1093/jb/mvm020
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© 2006 The Japanese Biochemical Society

The C-terminal tail of mitochondrial transcription factor A markedly strengthens its general binding to DNA

Kippei Ohgaki1, Tomotake Kanki1, Atsushi Fukuoh1, Hironori Kurisaki1, Yoshimasa Aoki1, Masaki Ikeuchi1,2, Sang Ho Kim1,3, Naotaka Hamasaki1 and Dongchon Kang1

1Department of Clinical Chemistry and Laboratory Medicine, Kyushu University, Graduate School of Medical Sciences, Fukuoka Japan
2Department of Cardiovascular Medicine, Kyushu University Graduate School of Medical Sciences, Fukuoka Japan
3Department of Biology Education, Daegu University, Kyungsan, Korea.

Dongchon Kang Department of Clinical Chemistry and Laboratory Medicine, Kyushu University Graduate School of Medical Sciences, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582 Japan, Tel: +81-92-642-5749 Fax: +81-92-642-5772 E-mail: kang{at}mailserver.med.kyushu-u.ac.jp

Received November 27, 2006; Accepted November 29, 2006


  Abstract

Mitochondrial transcription factor A (TFAM) contains a basic C-terminal tail which is essential for the promoter-specific transcription. TFAM is also a major component of a protein- mitochondrial DNA (mtDNA) complex, called nucleoid, as a nonspecific DNA-binding protein. However, little is known about a role of the C-tail in the nucleoid. Overexpression of full-length TFAM decreased the amount of a D-loop form of mtDNA in cells while overexpression of TFAM lacking its C-tail (TFAM-{Delta}C) did not, suggesting that the C-tail is involved in destabilization or formation of the D-loop. An mRNA for mtDNA-derived ND1 was hardly decreased in the former but rather decreased in the latter. Given that the D-loop formation is coupled with the transcription, the decrease in the D-loop is likely due to its destabilization. The recombinant full-length TFAM much strongly unwound DNA than TFAM-{Delta}C, which is consistent with the above idea because D-loop is resolved by unwinding of the supercoiling state. Notably, truncation of the C-tail decreased DNA-binding activity of TFAM by three orders of magnitude. Thus, the C-terminal tail of TFAM is important for the strong general binding to mtDNA. This strong DNA-binding conferred by the C-tail may play an important role in the nucleoid structure.

Key Words: D-loop, mitochondrial DNA, nucleoid, TFAM, transcription


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