TY - JOUR
T1 - Phosphorylation of PACSIN2 by protein kinase C triggers the removal of caveolae from the plasma membrane
AU - Senju, Yosuke
AU - Rosenbaum, Eva
AU - Shah, Claudio
AU - Hamada-Nakahara, Sayaka
AU - Itoh, Yuzuru
AU - Yamamoto, Kimiko
AU - Hanawa-Suetsugu, Kyoko
AU - Daumke, Oliver
AU - Suetsugu, Shiro
N1 - Funding Information:
This work was supported by grants from the Funding Program for Next Generation World-Leading Researchers (NEXT program); Japan Society for the Promotion of Science (JSPS) KAKENHI [grant number 26291037, 15H01641]; Astellas Foundation for Research on Metabolic Disorders to S.S.; European Research Council (ERC) consolidator [grant number ERC-2013-CoG-616024 to O.D.]; and a grant of the German Research Foundation [grant number SFB958/A12].
Publisher Copyright:
© 2015.
PY - 2015
Y1 - 2015
N2 - PACSIN2, a membrane-sculpting BAR domain protein, localizes to caveolae. Here, we found that protein kinase C (PKC) phosphorylates PACSIN2 at serine 313, thereby decreasing its membrane binding and tubulation capacities. Concomitantly, phosphorylation decreased the time span for which caveolae could be tracked at the plasma membrane (the 'tracking duration'). Analyses of the phospho-mimetic S313E mutant suggested that PACSIN2 phosphorylation was sufficient to reduce caveolartracking durations. Both hypotonic treatment and isotonic druginduced PKC activation increased PACSIN2 phosphorylation at serine 313 and shortened caveolar-tracking durations. Caveolartracking durations were also reduced upon the expression of other membrane-binding-deficient PACSIN2 mutants or upon RNA interference (RNAi)-mediated PACSIN2 depletion, pointing to a role for PACSIN2 levels in modulating the lifetime of caveolae. Interestingly, the decrease in membrane-bound PACSIN2 was inversely correlated with the recruitment and activity of dynamin 2, a GTPase that mediates membrane scission. Furthermore, expression of EHD2, which stabilizes caveolae and binds to PACSIN2, restored the tracking durations of cells with reduced PACSIN2 levels. These findings suggest that the PACSIN2 phosphorylation decreases its membrane-binding activity, thereby decreasing its stabilizing effect on caveolae and triggering dynaminmediated removal of caveolae.
AB - PACSIN2, a membrane-sculpting BAR domain protein, localizes to caveolae. Here, we found that protein kinase C (PKC) phosphorylates PACSIN2 at serine 313, thereby decreasing its membrane binding and tubulation capacities. Concomitantly, phosphorylation decreased the time span for which caveolae could be tracked at the plasma membrane (the 'tracking duration'). Analyses of the phospho-mimetic S313E mutant suggested that PACSIN2 phosphorylation was sufficient to reduce caveolartracking durations. Both hypotonic treatment and isotonic druginduced PKC activation increased PACSIN2 phosphorylation at serine 313 and shortened caveolar-tracking durations. Caveolartracking durations were also reduced upon the expression of other membrane-binding-deficient PACSIN2 mutants or upon RNA interference (RNAi)-mediated PACSIN2 depletion, pointing to a role for PACSIN2 levels in modulating the lifetime of caveolae. Interestingly, the decrease in membrane-bound PACSIN2 was inversely correlated with the recruitment and activity of dynamin 2, a GTPase that mediates membrane scission. Furthermore, expression of EHD2, which stabilizes caveolae and binds to PACSIN2, restored the tracking durations of cells with reduced PACSIN2 levels. These findings suggest that the PACSIN2 phosphorylation decreases its membrane-binding activity, thereby decreasing its stabilizing effect on caveolae and triggering dynaminmediated removal of caveolae.
KW - BAR domain
KW - Caveolae
KW - Mechanical stress
KW - Phosphorylation
KW - Protein kinase C
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U2 - 10.1242/jcs.167775
DO - 10.1242/jcs.167775
M3 - Article
C2 - 26092940
AN - SCOPUS:84974539428
VL - 128
SP - 2766
EP - 2780
JO - The Quarterly journal of microscopical science
JF - The Quarterly journal of microscopical science
SN - 0021-9533
IS - 15
ER -