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Molecular and Cellular Biology, October 1998, p. 5868-5879, Vol. 18, No. 10
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

The Deafness-Associated Mitochondrial DNA Mutation at Position 7445, Which Affects tRNA^Ser(UCN) Precursor Processing, Has Long-Range Effects on NADH Dehydrogenase Subunit ND6 Gene Expression

Min-Xin Guan,¹ José Antonio Enriquez,^1, Nathan Fischel-Ghodsian,² Ram S. Puranam,^1, Catherine P. Lin,¹ Marion A. Maw,³ and Giuseppe Attardi^1,*

Division of Biology, California Institute of Technology, Pasadena, California 91125¹; Ahmanson Department of Pediatrics, Steven Spielberg Pediatric Research Center, Cedars-Sinai Medical Center, Los Angeles, California 90048²; and Department of Biochemistry, University of Otago, Dunedin, New Zealand³

Received 30 April 1998/Accepted 11 July 1998

The pathogenetic mechanism of the deafness-associated mitochondrial DNA (mtDNA) T7445C mutation has been investigated in several lymphoblastoid cell lines from members of a New Zealand pedigree exhibiting the mutation in homoplasmic form and from control individuals. We show here that the mutation flanks the 3' end of the tRNA^Ser(UCN) gene sequence and affects the rate but not the sites of processing of the tRNA precursor. This causes an average reduction of ~70% in the tRNA^Ser(UCN) level and a decrease of ~45% in protein synthesis rate in the cell lines analyzed. The data show a sharp threshold in the capacity of tRNA^Ser(UCN) to support the wild-type protein synthesis rate, which corresponds to ~40% of the control level of this tRNA. Strikingly, a 7445 mutation-associated marked reduction has been observed in the level of the mRNA for the NADH dehydrogenase (complex I) ND6 subunit gene, which is located ~7 kbp upstream and is cotranscribed with the tRNA^Ser(UCN) gene, with strong evidence pointing to a mechanistic link with the tRNA precursor processing defect. Such reduction significantly affects the rate of synthesis of the ND6 subunit and plays a determinant role in the deafness-associated respiratory phenotype of the mutant cell lines. In particular, it accounts for their specific, very significant decrease in glutamate- or malate-dependent O₂ consumption. Furthermore, several homoplasmic mtDNA mutations affecting subunits of NADH dehydrogenase may play a synergistic role in the establishment of the respiratory phenotype of the mutant cells.

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^* Corresponding author. Mailing address: Division of Biology, California Institute of Technology, Pasadena, CA 91125. Phone: (626) 395-4930. Fax: (626) 449-0756. E-mail: attardig{at}seqaxp.bio.caltech.edu.

Present address: Departamento de Bioquímica, Universidad de Zaragoza, 50013 Zaragoza, Spain.

Present address: Department of Medicine, Duke University Medical Center, Durham, NC 27710.

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Molecular and Cellular Biology, October 1998, p. 5868-5879, Vol. 18, No. 10
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

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