Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 2766-2780 |
| Number of pages | 15 |
| Journal | Journal of cell science |
| Volume | 128 |
| Issue number | 15 |
| DOIs | |
| Publication status | Published - Jan 1 2015 |
| Externally published | Yes |
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Keywords
- BAR domain
- Caveolae
- Mechanical stress
- Phosphorylation
- Protein kinase C
ASJC Scopus subject areas
- Cell Biology
Cite this
Phosphorylation of PACSIN2 by protein kinase C triggers the removal of caveolae from the plasma membrane. / Senju, Yosuke; Rosenbaum, Eva; Shah, Claudio; Hamada-Nakahara, Sayaka; Itoh, Yuzuru; Yamamoto, Kimiko; Hanawa-Suetsugu, Kyoko; Daumke, Oliver; Suetsugu, Shiro.
In: Journal of cell science, Vol. 128, No. 15, 01.01.2015, p. 2766-2780.Research output: Contribution to journal › Article
}
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
PY - 2015/1/1
Y1 - 2015/1/1
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
UR - http://www.scopus.com/inward/record.url?scp=84974539428&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84974539428&partnerID=8YFLogxK
U2 - 10.1242/jcs.167775
DO - 10.1242/jcs.167775
M3 - Article
C2 - 26092940
AN - SCOPUS:84974539428
VL - 128
SP - 2766
EP - 2780
JO - Journal of Cell Science
JF - Journal of Cell Science
SN - 0021-9533
IS - 15
ER -