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PMID: 15703217 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Phosphorylation controls CLIMP-63-mediated anchoring of the endoplasmic reticulum to microtubules.

Molecular biology of the cell ·Vol. 16 ·No. 4 ·2005-04-00 ·Pages 1928-37

Vedrenne C, Klopfenstein DR, Hauri HP

Abstract

The microtubule-binding 63-kDa cytoskeleton-linking membrane protein (CLIMP-63) is an integral membrane protein that links the endoplasmic reticulum (ER) to microtubules. Here, we tested whether this interaction is regulated by phosphorylation. Metabolic labeling with (32)P showed that CLIMP-63 is a phosphoprotein with increased phosphorylation during mitosis. CLIMP-63 of mitotic cells is unable to bind to microtubules in vitro. Mitotic phosphorylation can be prevented by mutation of serines 3, 17, and 19 in the cytoplasmic domain of CLIMP-63. When these residues are mutated to glutamic acid, and hence mimic mitotic phosphorylation, CLIMP-63 does no longer bind to microtubules in vitro. Overexpression of the phospho-mimicking mitotic form of CLIMP-63 in interphase cells leads to a collapse of the ER around the nucleus, leaving the microtubular network intact. The results suggest that CLIMP-63-mediated stable anchoring of the ER to microtubules is required to maintain the spatial distribution of the ER during interphase and that this interaction is abolished by phosphorylation of CLIMP-63 during mitosis.

MeSH Terms
Amino Acid Sequence Animals Cell Line Chlorocebus aethiops Endoplasmic Reticulum/metabolism Humans Membrane Proteins/chemistry,genetics,metabolism Microtubules/metabolism Mitosis Molecular Sequence Data Mutation/genetics Phosphorylation Phosphoserine/metabolism
Chemicals
CKAP4 protein, human Membrane Proteins Phosphoserine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Vedrenne Cécile
Biozentrum, University of Basel, Switzerland.
Klopfenstein Dieter R
Hauri Hans-Peter
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e04-07-0554
Published
2005-04-00
Epub
2005-00-09
Pages
1928-37
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1073672
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.e04-07-0554 PMC Full Text View journal details
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