The Study Of Pink1-mediated Phosphorylation To Influence Mitophagy

Parkinson's Disease(PD)is a neurodegenerative disease caused by multiple factors.Studies indicate that the main cause of PD is the degeneration and loss of dopamine neurons in the dense part of the substantia nigra.Since the 19th century,humans have had 200 years of research and clinical treat ment of PD,but the specific etiology and pathogenesis of PD have not been proved yet.With the development of molecular biology technology,the molecular mechanism of PD pathogenesis has become a clue for researchers.Mitophagy is a fundamental process in which cells selectively remove unnecessary or damaged mitochondria,which is an important part of the mitochondrial quality Control.Current research suggests that mitophagy can trigger a series of neurodegenerative diseases.A PD associated gene which is named PARK6,its abnormal expression of PINK1 kinase which coded by PARK6 can lead to dysfunction such as mitophagy,which eventually causes PD symptoms.At present,the relationship between the homeostasis of mitophagy and PD is still unknown.Following the clue of mitophagy mediated by Pinkl,we have used Co-IP and MS analysis to screen out a kind of mitochondrial targeting protein,JK2F.We also have used Ser/Thr MS analysis and in vitro kinase assay to determine Pinkl could phosphorylate the S412 sites of JK2F(JK2F-S412)exactlly.To this end,we have leaved two questions:What the effect of Pinkl/JK2F in the regulation of mitophagy?What is the role of JK2F in PD,These are the main contents of this research in Drosophila and Hela cells.Our results showed that the combination of PINK1 and JK2F in vivo is not affected by mitochondria targeting sequences by using Co-IP.The physiological function of the test revealed that overexpression of JK2F could restore mitophagy in the background of PINK1 defects.In order to explore the regulation mechanism,the phosphorylation of JK2F-Ser412 is medied by Pink1-dependence manner through(p-s412)JK2F antibody in WB detection.The effect of JK2F's phosphorylation on mitophagy was detected at protein and cell level,the results showed the S412A state could promote the process of mitophagy,in contract,S412D state would inhibit this process.At the same time,we found that the CCCP induced mitophagy in different cell lines.The phosphorylation of JK2F began to occur in the early stage and continued to increase after the 6-8h began to decay.This trend is in sync with PINK1's ability to induce mitophagy.In addition,it was found that the variation of JK2F abundance could affect the avg.area of mitochondria.And it was also found the degree of JK2F transport to mitochondria was associate with the aging in Drosophila background studies.Here we have preliminarily revealed that Pinkl through mediated JK2F phosphorylation to perform bidirectional autophagy regulation on mitochondria,in order to prevent elimination of Mitochondria from the excessive autophagy to destroy the homeostasis of the intracellular environment.Moreover,the effect of JK2F on the mitochondria quality is to ensure the normal function of the cells.Our results provides a new clue for PD studies with the impaired functional mitochondrial.