NDLI: M Phase-specific Expression and Phosphorylation-dependent Ubiquitination of the ClC-2 Channel

Content Provider	Journal of Biological Chemistry (JBC)
Author	Zheng, Ya-Juan Furukawa, Tetsushi Ogura, Takehiko Tajimi, Kimitaka Inagaki, Nobuya
Abstract	Cl− channel activities vary during the cell cycle and are thought to play various roles including regulation of cell volume. We have shown previously that ClC-2 channels are directly phosphorylated and functionally regulated by the M phase-specific cyclin-dependent kinase p34cdc2/cyclin B. We investigate here to determine whether the expression levels of ClC-2 channel protein vary during the cell cycle. Immunoblot and immunocytochemical analyses of cells cycle-synchronized by serum depletion/replenishment reveal that ClC-2 channel protein is expressed predominantly at M phase in cells with two nuclei and a clear constriction ring, whereas RNA blot analysis shows that ClC-2 mRNA expression does not change during the cell cycle. Ubiquitin assays reveal that the ClC-2 channels are ubiquitinated at M phase, whereas the magnitude of ubiquitination is suppressed by incubation with olomoucine, an inhibitor of p34cdc2/cyclin B, and it is almost completely abolished in ClC-2 channels having an S632A mutation, which cannot be phosphorylated by p34cdc2/cyclin B, indicating that ubiquitination of ClC-2 channels requires phosphorylation by M phase-specific p34cdc2/cyclin B. Regulation at the post-transcriptional level, including phosphorylation-dependent ubiquitination, may contribute to M phase-specific expression of ClC-2 channels. Cell cycle-dependent regulation of expression at the protein level in addition to the regulation of function suggests that the ClC-2 channel plays a physiological role in the cell cycle.
Related Links	http://www.jbc.org/content/277/35/32268.abstract
Ending Page	32273
Starting Page	32268
Page Count	6
File Format	HTM / HTML PDF
ISSN	00219258
Journal	Journal of Biological Chemistry (JBC)
Issue Number	35
Volume Number	277
DOI	10.1074/jbc.M202105200
e-ISSN	1083351X
Language	English
Publisher	American Society for Biochemistry and Molecular Biology
Publisher Date	2002-08-30
Access Restriction	Open
Subject Keyword	Hemagglutinin (HA) Fetal calf serum (FCS) Benzyloxycarbonyl-leucyl-leucyl-leucinal (zLLLal) Dimethyl sulfoxide (Me2SO) MOLECULAR BASIS OF CELL AND DEVELOPMENTAL BIOLOGY
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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