a flow cytometry-based screen of nuclear envelope transmembrane proteins identifies net4tmem53 as involved in stress-dependent cell cycle withdrawal屏幕流cytometry-based核膜的跨膜蛋白识别net4tmem53参与细胞周期stress-dependent撤军.pdfVIP

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a flow cytometry-based screen of nuclear envelope transmembrane proteins identifies net4tmem53 as involved in stress-dependent cell cycle withdrawal屏幕流cytometry-based核膜的跨膜蛋白识别net4tmem53参与细胞周期stress-dependent撤军.pdf

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a flow cytometry-based screen of nuclear envelope transmembrane proteins identifies net4tmem53 as involved in stress-dependent cell cycle withdrawal屏幕流cytometry-based核膜的跨膜蛋白识别net4tmem53参与细胞周期stress-dependent撤军

A Flow Cytometry-Based Screen of Nuclear Envelope Transmembrane Proteins Identifies NET4/Tmem53 as Involved in Stress-Dependent Cell Cycle Withdrawal 1. 1. 2 1 3 Nadia Korfali , Vlastimil Srsen , Martin Waterfall , Dzmitry G. Batrakou , Vanja Pekovic , 3 1 Christopher J. Hutchison , Eric C. Schirmer * 1The Wellcome Trust Centre for Cell Biology and Institute of Cell Biology, University of Edinburgh, Edinburgh, United Kingdom, 2 Institute of Immunology and Infection Research, University of Edinburgh, Edinburgh, United Kingdom, 3 School of Biological and Biomedical Sciences, Durham University, Durham, United Kingdom Abstract Disruption of cell cycle regulation is one mechanism proposed for how nuclear envelope protein mutation can cause disease. Thus far only a few nuclear envelope proteins have been tested/found to affect cell cycle progression: to identify others, 39 novel nuclear envelope transmembrane proteins were screened for their ability to alter flow cytometry cell cycle/ DNA content profiles when exogenously expressed. Eight had notable effects with seven increasing and one decreasing the 4N:2N ratio. We subsequently focused on NET4/Tmem53 that lost its effects in p532/ 2 cells and retinoblastoma protein- deficient cells. NET4/TMEM53 knockdown by siRNA altered flow cytometry cell cycle/DNA content profiles in a similar way as overexpression. NET4/TMEM53 knockdown did not affect total retinoblastoma protein levels, unlike nuclear envelope- associated proteins Lamin A and LAP2a. However, a decrease in phosphorylated retinoblastoma protein was observed along with a doubling of p53 levels and a 7-fold increase in p21. Consequently ce