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The Effects of PTPA on PP2A_c Y307 Phosphorylation Level and the Underlying Mechanisms

Author: NieYunJuan
Tutor: ZhouXinWen
School: Huazhong University of Science and Technology
Course: Pathology and Pathophysiology
Keywords: Protein Phosphatase Activity of factor Protein phosphatase -2A Tyrosine 307 Protein tyrosine phosphatase 1B tau protein Phosphorylation
CLC: R749.16
Type: Master's thesis
Year: 2011
Downloads: 16
Quote: 0
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Abstract


Alzheimer's disease (Alzheimer's disease, AD) is a progressive memory loss and cognitive impairment neurodegenerative diseases, two major pathological changes characteristic of AD brain of patients with the formation of a large number of neurofibrillary tangle neurons Results (neurofibrillary tangles, NFT), and the large number of cell senile plaques (senile plaques, SP) deposition. Wherein the main component of the neurofibrillary tangles are abnormally phosphorylated tau protein. The human body has 107 tyrosine phosphatase PP2A belonging to the PPP family, play an important role in phosphatase in AD development process affects tau phosphorylation. PP2A catalytic subunit activity regulation: change PP2AC expression levels as well as the translation of PP2AC modification - Leu309 methylation and Tyr307 phosphorylation. PTPA is a ubiquitous protein in mammalian tissues and cells, PTPA is able to enhance the activity of PP2A phosphatase, by changing PP2AC spatial conformation and increase the level of methylation of the the Leu309 bit point but PP2AC Tyr307 phosphorylation sites level whether there is a role, it is not clear. Objective: To study the impact of PTPA PP2AC Tyr307 phosphorylation levels and its possible mechanism. Materials and Methods: build the PTPA wild-type plasmid (wt-PTPA) and specificity for the the PTPA gene sequences of RNA interference the plasmid the pSUPER-siPTPA (siPTPA) respectively transfected into cell lines of HEK293/tau, the protein immunoblotting tau protein Members phosphorylation levels, detect tau phosphorylation kinases, PP2AC protein the, PP2AC demethylation, phosphorylation levels of change. Transfected PP2A activity kits for detecting changes in PP2A activity; cells co-localization observed by immunofluorescence the PTPA and tau phosphorylation sites; to detect changes in PTP1B transcriptional level by RT-PCR method. Results: (1) wtPTPA effectively increase the expression of the PTPA, pSUP-siPTPA effectively reduce the expression of the PTPA; (2) 293/tau cells, over-expression The PTPA can reduce the level of tau phosphorylation, knockout PTPA tau protein phosphorylation increase in the level; (3) regulation of PTPA expression levels change PP2A activity, but no effect on GSK-3β; (4) OA reversible reduction of the level of tau phosphorylation after PTPA turned his expression; (5) overexpression PTPA can reduce of PP2AC Y307 phosphorylation; PTPA knockout, after PP2ACY307 phosphorylation level rise; (6) overexpression PTPA can increase the phosphatase PTP1B's PP2AC Y307 loci transcription and protein expression level; PTPA knockout after transcription and protein expression of PTP1B levels decreased, while the other causes change PP2AC Y307 kinase not changed. Conclusion: the PTPA regulation PTP1B mRNA and protein levels and to tyrosine phosphorylation affect PP2AC307 sites.

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