Abstract
RAS association domain family 4 (RASSF4) is involved in tumorigenesis and regulation of the Hippo pathway. In this study, we identify new functional roles of RASSF4. First, we discovered that RASSF4 regulates store-operated Ca2+ entry (SOCE), a fundamental Ca2+ signaling mechanism, by affecting the translocation of the endoplasmic reticulum (ER) Ca2+ sensor stromal interaction molecule 1 (STIM1) to ER-plasma membrane (PM) junctions. It was further revealed that RASSF4 regulates the formation of ER-PM junctions and the ER-PM tethering function of extended synaptotagmins E-Syt2 and E-Syt3. Moreover, steady-state PM phosphatidylinositol 4,5-bisphosphate (PI[4,5]P2) levels, important for localization of STIM1 and E-Syts at ER-PM junctions, were reduced in RASSF4-knockdown cells. Furthermore, we demonstrated that RASSF4 interacts with and regulates the activity of adenosine diphosphate ribosylation factor 6 (ARF6), a small G protein and upstream regulator of type I phosphatidylinositol phosphate kinases (PIP5Ks) and PM PI(4,5)P2 levels. Overall, our study suggests that RASSF4 controls SOCE and ER-PM junctions through ARF6-dependent regulation of PM PI(4,5)P2 levels, pivotal for a variety of physiological processes.
© 2017 Chen et al.
MeSH terms
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ADP-Ribosylation Factor 6
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ADP-Ribosylation Factors / genetics
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ADP-Ribosylation Factors / metabolism
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Calcium Signaling*
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Cell Membrane / metabolism*
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Endoplasmic Reticulum / metabolism*
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Female
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HeLa Cells
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Humans
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Microscopy, Fluorescence
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Microscopy, Video
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Neoplasm Proteins / genetics
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Neoplasm Proteins / metabolism*
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ORAI1 Protein / metabolism*
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Phosphatidylinositol 4,5-Diphosphate / metabolism*
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Phosphotransferases (Alcohol Group Acceptor) / genetics
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Phosphotransferases (Alcohol Group Acceptor) / metabolism
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Protein Transport
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RNA Interference
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Stromal Interaction Molecule 1 / genetics
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Stromal Interaction Molecule 1 / metabolism*
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Synaptotagmin II / genetics
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Synaptotagmin II / metabolism
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Synaptotagmins / genetics
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Synaptotagmins / metabolism
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Time Factors
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Time-Lapse Imaging
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Transfection
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Tumor Suppressor Proteins / genetics
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Tumor Suppressor Proteins / metabolism*
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Uterine Cervical Neoplasms / genetics
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Uterine Cervical Neoplasms / metabolism
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Uterine Cervical Neoplasms / pathology
Substances
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ADP-Ribosylation Factor 6
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Neoplasm Proteins
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ORAI1 Protein
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ORAI1 protein, human
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Phosphatidylinositol 4,5-Diphosphate
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RASSF4 protein, human
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STIM1 protein, human
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SYT2 protein, human
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SYT3 protein, human
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Stromal Interaction Molecule 1
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Synaptotagmin II
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Tumor Suppressor Proteins
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Synaptotagmins
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Phosphotransferases (Alcohol Group Acceptor)
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1-phosphatidylinositol-4-phosphate 5-kinase
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ADP-Ribosylation Factors
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ARF6 protein, human