[基础医学]MIDAS.pptVIP

* * DNA sequencing distinguished the gene corresponding to the third band in Fig. 1A from the genes which have been so far identi?ed to be involved in mitochondrial biogenesis, gene expression and metabolism. The authors named this gene MIDAS (mitochondrial DNA absence sensitive factor),because the gene product was expressed in response to mtDNA depletion. Interestingly, the nucleotide sequence was identical to GPP34 whose product has been identi?ed as a Golgi protein of unknown function(s). * Results MIDAS/GPP34 was expressed more abundantly in EB8 than in Ft2-11. Therefore, the authors focused on the function of MIDAS/GPP34. * Results 2.Speci?c expression of MIDAS/GPP34 in muscle ?bers with abnormal mitochondria The authors examined the expression of MIDAS/GPP34 in muscle ?bers of patients with mitochondrial diseases CPEO and MELAS. (CPEO 慢性进行性眼外肌麻痹、MELAS线粒体脑肌病伴高乳酸血症和卒中样发作). Accumulations of abnormal mitochondria are detected as ragged-red ?bers(破碎样红纤维) and high succinate dehydrogenase (SDH琥珀酸脱氢酶) ?bers, with mutant mtDNA dominating in a mosaic manner . * No MIDAS-positive ?bers were detected in normal muscle, whereas MIDAS-positive ?bers were detected in the muscle sections of affected patients. Thus, MIDAS is expressed in response to mitochondrial dysfunction in muscle with mitochondrial diseases. (Fig. 2) Muscle sections from a normal subject and patients with CPEO or MELAS were stained for activity of SDH and cytochrome c oxidase (COX) and with anti-MIDAS antibody. * Results 3.Subcellular distribution of MIDAS/GPP34 ①We have already known that GPP34 was isolated as a Golgi peripheral membrane protein in a Golgi proteomics study . To verify the distribution of MIDAS in mitochondria, the authors immunostained HeLa cells with an affinity-purified polyclonal antibody against the MIDAS protein. Immunostaining of HeLa cells revealed that MIDAS mainly colocalized with MitoTracker Red (Fig. 3A). * With careful observation, the authors found MIDAS in an addi