Defining the Importance of Mitochondrial Gene Defects in Maternally Inherited Diabetes by Sequencing the Entire Mitochondrial Genome

Defining the Importance of Mitochondrial Gene Defects in Maternally Inherited Diabetes by Sequencing the Entire Mitochondrial Genome Alan T.W. Choo-Kang 1 , Stephen Lynn 1 , Geoffrey A. Taylor 2 , Mark E. Daly 1 , Sarbpreet S. Sihota 1 , Teressa M. Wardell 2 , Patrick F. Chinnery 2 , Douglass M. Turnbull 2 and Mark Walker 1 1 School of Clinical Medical Sciences, University of Newcastle upon Tyne, U.K. 2 School of Clinical Neurosciences, University of Newcastle upon Tyne, U.K. Abstract For any mitochondrial DNA (mtDNA) mutation, the ratio of mutant to wild-type mtDNA (% heteroplasmy) varies across tissues, with low levels in leukocytes and high levels in postmitotic tissues (e.g., skeletal muscle). Direct sequencing is the gold-standard method used to detect novel mutations, but can only reliably detect % heteroplasmy >25%, which is rare in leukocytes. Therefore, we investigated the role of mtDNA defects in maternally inherited diabetes by first screening for the A3243G tRNA Leu(UUR) mutation by restriction assay, followed by sequencing of the entire mitochondrial genome using skeletal muscle derived mtDNA. A total of 28 patients had maternally inherited diabetes either alone (group 1, n = 17) or with one or more additional features of mitochondrial disease, including bilateral sensori-neural deafness and neuromuscular disease (group 2, n = 11). Three patients (all from group 2) carried the A3243G mutation. Skeletal muscle mtDNA from eight group 1 patients and six more group 2 patients was sequenced. No pathogenic mutations were found in the group 1 patients, while two patients from group 2 had mutations at positions 12258 and 14709 in the tRNA serine and glutamic acid genes, respectively. We conclude, therefore, that screening for mtDNA mutations should be considered in patients with maternally inherited diabetes, but only when additional features of mitochondrial disease are present. Footnotes Address correspondence and reprint requests to Dr. Mark Walker, School of Clinical Medical Sciences, 4th William Leech Block, The Medical School, Framlington Place, Newcastle upon Tyne, U.K., NE2 4HH. E-mail: mark.walker{at}ncl.ac.uk . Received for publication 24 January 2002 and accepted in revised form 3 April 2002. S.L. is currently affiliated with the Department of Pediatrics, University of California, San Francisco, California. % heteroplasmy, ratio of mutant to wild-type mtDNA; ICA, islet cell antibody; MIDD, maternally inherited diabetes and deafness; mtDNA,