PMID- 25861040
OWN - NLM
STAT- MEDLINE
DCOM- 20160418
LR  - 20181113
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 10
IP  - 4
DP  - 2015
TI  - Filamin-a increases the stability and plasma membrane expression of polycystin-2.
PG  - e0123018
LID - 10.1371/journal.pone.0123018 [doi]
LID - e0123018
AB  - Polycystin-2 (PC2), encoded by the PKD2 gene, is mutated in ~15% of autosomal 
      dominant polycystic kidney disease. Filamins are actin-binding proteins 
      implicated in scaffolding and membrane stabilization. Here we studied the effects 
      of filamin on PC2 stability using filamin-deficient human melanoma M2, filamin-A 
      (FLNA)-replete A7, HEK293 and IMCD cells together with FLNA siRNA/shRNA knockdown 
      (KD). We found that the presence of FLNA is associated with higher total and 
      plasma membrane PC2 protein expression. Western blotting analysis in combination 
      with FLNA KD showed that FLNA in A7 cells represses PC2 degradation, prolonging 
      the half-life from 2.3 to 4.4 hours. By co-immunoprecipitation and Far Western 
      blotting we found that the FLNA C-terminus (FLNAC) reduces the FLNA-PC2 binding 
      and PC2 expression, presumably through competing with FLNA for binding PC2. We 
      further found that FLNA mediates PC2 binding with actin through forming complex 
      PC2-FLNA-actin. FLNAC acted as a blocking peptide and disrupted the link of PC2 
      with actin through disrupting the PC2-FLNA-actin complex. Finally, we 
      demonstrated that the physical interaction of PC2-FLNA is Ca-dependent. Taken 
      together, our current study indicates that FLNA anchors PC2 to the actin 
      cytoskeleton through complex PC2-FLNA-actin to reduce degradation and increase 
      stability, and possibly regulate PC2 function in a Ca-dependent manner.
FAU - Wang, Qian
AU  - Wang Q
AD  - Membrane Protein Disease Research Group, Department of Physiology, Faculty of 
      Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada.
FAU - Zheng, Wang
AU  - Zheng W
AD  - Membrane Protein Disease Research Group, Department of Physiology, Faculty of 
      Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada.
FAU - Wang, Zuocheng
AU  - Wang Z
AD  - Membrane Protein Disease Research Group, Department of Physiology, Faculty of 
      Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada.
FAU - Yang, JungWoo
AU  - Yang J
AD  - Membrane Protein Disease Research Group, Department of Physiology, Faculty of 
      Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada.
FAU - Hussein, Shaimaa
AU  - Hussein S
AD  - Membrane Protein Disease Research Group, Department of Physiology, Faculty of 
      Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada.
FAU - Tang, Jingfeng
AU  - Tang J
AD  - Membrane Protein Disease and Cancer Research Center, Hubei University of 
      Technology, Wuhan, China; Membrane Protein Disease Research Group, Department of 
      Physiology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, 
      Alberta, Canada.
FAU - Chen, Xing-Zhen
AU  - Chen XZ
AD  - Membrane Protein Disease and Cancer Research Center, Hubei University of 
      Technology, Wuhan, China; Membrane Protein Disease Research Group, Department of 
      Physiology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, 
      Alberta, Canada.
LA  - eng
GR  - MOP 89977/Canadian Institutes of Health Research/Canada
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150410
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Filamins)
RN  - 0 (Multiprotein Complexes)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (TRPP Cation Channels)
RN  - 0 (polycystic kidney disease 2 protein)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Actin Cytoskeleton/metabolism
MH  - Animals
MH  - Calcium/metabolism
MH  - Cell Line
MH  - Cell Membrane/metabolism
MH  - Filamins/antagonists & inhibitors/genetics/*metabolism
MH  - Gene Knockdown Techniques
MH  - Green Fluorescent Proteins/genetics/metabolism
MH  - HEK293 Cells
MH  - Humans
MH  - Mice
MH  - Multiprotein Complexes/chemistry/genetics/metabolism
MH  - Protein Interaction Domains and Motifs
MH  - Protein Stability
MH  - Proteolysis
MH  - Recombinant Fusion Proteins/chemistry/genetics/metabolism
MH  - TRPP Cation Channels/chemistry/genetics/*metabolism
PMC - PMC4393133
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2015/04/11 06:00
MHDA- 2016/04/19 06:00
PMCR- 2015/04/10
CRDT- 2015/04/11 06:00
PHST- 2014/10/29 00:00 [received]
PHST- 2015/02/26 00:00 [accepted]
PHST- 2015/04/11 06:00 [entrez]
PHST- 2015/04/11 06:00 [pubmed]
PHST- 2016/04/19 06:00 [medline]
PHST- 2015/04/10 00:00 [pmc-release]
AID - PONE-D-14-48717 [pii]
AID - 10.1371/journal.pone.0123018 [doi]
PST - epublish
SO  - PLoS One. 2015 Apr 10;10(4):e0123018. doi: 10.1371/journal.pone.0123018. 
      eCollection 2015.