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Am J Physiol Endocrinol Metab (November 6, 2001). doi:10.1152/ajpendo.00255.2001
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Articles in PresS, published online ahead of print November 5, 2001
Am J Physiol Endocrinol Metab, 10.1152/ajpendo.00255.2001
Submitted on June 13, 2001
Accepted on October 31, 2001

Reduced activity of mtTFA decreases the transcription in mitochondria isolated from diabetic rat heart

Akio Kanazawa1, Yoshihiko Nishio2, Atsunori Kashiwagi2*, Hidetoshi Inagaki3, Ryuichi Kikkawa2, and Kihachiro Horiike1

1 Department of Biochemistry, Shiga University of Medical Science, Otsu, Shiga, Japan
2 Third Department of Medicine, Shiga University of Medical Science, Otsu, Shiga, Japan
3 Molecular Neurophysiology Group, National Institute of Advanced Industrial Science and Technology, Tukuba, Ibaragi, Japan

* To whom correspondence should be addressed. E-mail: kasiwagi{at}belle.shiga-med.ac.jp.

To evaluate abnormalities in the mitochondrial transcription factor A (mtTFA) function as a cause of mitochondrial dysfunction in diabetes, we measured the mRNA contents of the proteins consisting of the mitochondrial respiratory chain as well as transcriptional and translational activities in the mitochondria isolated from controls and streptozotocin-induced diabetic rat hearts. Using Northern blot analysis, we found the 40% reduced mRNA contents of mitochondrial-encoded cytochrome b and ATP synthase subunit 6 in diabetic rat hearts compared with control rats (P < 0.05). These abnormalities were completely recovered by insulin treatment. Furthermore, the mitochondrial activities of transcription and translation were decreased significantly in mitochondria isolated from diabetic rats by 60% (P < 0.01) and by 71% (P < 0.01), respectively compared with control rats. The insulin treatment also completely normalized these abnormalities in diabetic rats. Consistently, gel retardation assay showed a reduced binding of mtTFA to the D-loop of mitochondrial DNA in diabetic rats, although there was no difference in the mtTFA mRNA and protein contents between the two groups. Based on these findings, a reduced binding activity of mtTFA to the D-loop region in the hearts of diabetic rats may contribute to the decreased mitochondrial protein synthesis.




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