The effects mutant DJ-1 and sumoylation of DJ-1 protein on DJ-1 mitochondrial localization / 中风与神经疾病杂志

ABSTRACT

@#Objective To investigate the effects of DJ-1 A39S mutation in Parkinson’s disease（PD）and SUMO modification of DJ-1 on DJ-1 mitochondrial localization. Methods Five COS-7 cell lines that can stably express wild type，A39S mutant，K130R mutant，A39S K130R double mutant DJ-1 protein and SUMO-1 protein were constructed. The transfection results were detected by Western blot，double enzyme digestion and PCR sequencing. The expression and subcellular localization of DJ-1 protein were observed by Western blotting，confocal microscope and immunofluorescence.Results Immunofluorescence showed that DJ-1 protein was mainly distributed in the cytoplasm and also in the nucleus.Some wild-type and K130R DJ-1 proteins were distributed in mitochondria，most of A39S and A39S-K130R mutant DJ-1 proteins were transferred to mitochondria，and SUMO-1 was mainly distributed in the nucleus and around the nucleus.Immunofluorescence co localization showed that four DJ-1 proteins were colocalize with SUMO-1 protein. Conclusion The pathogenic mutation A39S of DJ-1 transferred DJ-1 protein from cytoplasm to mitochondria；DJ-1 and SUMO-1 have colocalization，and they may interact.mitochondrial localization.Methods Five COS-7 cell lines that can stably express wild type，A39S mutant，K130R mutant，A39S K130R double mutant DJ-1 protein and SUMO-1 protein were constructed. The transfection results were detected by Western blot，double enzyme digestion and PCR sequencing.The expression and subcellular localization of DJ-1 protein were observed by Western blotting，confocal microscope and immunofluorescence.Results Immunofluorescence showed that DJ-1 protein was mainly distributed in the cytoplasm and also in the nucleus.Some wild type and K130R DJ-1 proteins were distributed in mitochondria，most of A39S and A39S-K130R mutant DJ-1 proteins were transferred to mitochondria，and SUMO-1 was mainly distributed in the nucleus and around the nucleus.Immunofluorescence colocalization showed that four DJ-1 proteins were colocalize with SUMO-1 protein.Conclusion The pathogenic mutation A39S of DJ-1 transferred DJ-1 protein from cytoplasm to mitochondria；DJ-1 and SUMO-1 have colocalization，and they may interact.