Analysis of Mitochondrial DNA Sequences in Childhood Encephalomyopathies Reveals New Disease-Associated Variants 英文参考文献.docVIP

AnalysisofMitochondrialDNASequencesinChildhood EncephalomyopathiesRevealsNewDisease-Associated Variants AijazA.Wani1,SajadH.Ahanger1,SharmilaA.Bapat1,AshrafY.Rangrez1,NitinHingankar2,C.G.Suresh2,ShamaBarnabas2,MilindS.Patole1, YogeshS.Shouche1* 1NationalCentreforCellScience,Pune,India,2DivisionofBiochemicalSciences,NationalChemicalLaboratory,Pune,India Background. Mitochondrial encephalomyopathies are a heterogeneous group ofclinical disorders generally caused due to mutationsineithermitochondrialDNA(mtDNA)ornucleargenesencodingoxidativephosphorylation(OXPHOS).Weanalyzed the mtDNA sequences from a group of 23 pediatric patients with clinical and morphological features of mitochondrial encephalopathies and tried to establish a relationship of identified variants with the disease. Methodology/Principle Findings. Complete mitochondrial genomes were amplified by PCR and sequenced by automated DNA sequencing. Sequencing data was analyzed by SeqScape software and also confirmed by BLASTn program. Nucleotide sequences were comparedwiththerevisedCambridgereferencesequence(CRS)andsequencespresentinmitochondrialdatabases.Thedata obtained shows that a number of known and novel mtDNA variants were associated with the disease. Most of the non- synonymousvariantswereheteroplasmic(A4136G,A9194GandT11916A)suggestingtheirpossibilityofbeingpathogenicin nature.Someofthemissensevariantsalthoughhomoplasmicwereshowingchangesinhighlyconservedaminoacids(T3394C, T3866C, and G9804A) and were previously identified with diseased conditions. Similarly, two other variants found in tRNA genes(G5783AandC8309T)couldalterthesecondarystructureofCys-tRNAandLys-tRNA.Mostofthevariantsoccurredin singlecases;however,afewoccurredinmorethanonecase(e.g.G5783AandA10149T).ConclusionsandSignificance.The mtDNA variants identified in this study could be the possible cause ofmitochondrial encephalomyopathies with childhood onset in the patient group. Our study further strengthens the pathogenic score of known v