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PMID: 20466722 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Conserved motif of CDK5RAP2 mediates its localization to centrosomes and the Golgi complex.

The Journal of biological chemistry ·Vol. 285 ·No. 29 ·2010-07-16 ·Pages 22658-65

Wang Z, Wu T, Shi L, Zhang L, Zheng W, Qu JY, Niu R, Qi RZ

Abstract

As the primary microtubule-organizing centers, centrosomes require gamma-tubulin for microtubule nucleation and organization. Located in close vicinity to centrosomes, the Golgi complex is another microtubule-organizing organelle in interphase cells. CDK5RAP2 is a gamma-tubulin complex-binding protein and functions in gamma-tubulin attachment to centrosomes. In this study, we find that CDK5RAP2 localizes to the Golgi complex in an ATP- and centrosome-dependent manner and associates with Golgi membranes independently of microtubules. CDK5RAP2 contains a centrosome-targeting domain with its core region highly homologous to the Motif 2 (CM2) of centrosomin, a functionally related protein in Drosophila. This sequence, referred to as the CM2-like motif, is also conserved in related proteins in chicken and zebrafish. Therefore, CDK5RAP2 may undertake a conserved mechanism for centrosomal localization. Using a mutational approach, we demonstrate that the CM2-like motif plays a crucial role in the centrosomal and Golgi localization of CDK5RAP2. Furthermore, the CM2-like motif is essential for the association of the centrosome-targeting domain to pericentrin and AKAP450. The binding with pericentrin is required for the centrosomal and Golgi localization of CDK5RAP2, whereas the binding with AKAP450 is required for the Golgi localization. Although the CM2-like motif possesses the activity of Ca(2+)-independent calmodulin binding, binding of calmodulin to this sequence is dispensable for centrosomal and Golgi association. Altogether, CDK5RAP2 may represent a novel mechanism for centrosomal and Golgi localization.

MeSH Terms
Amino Acid Motifs Calmodulin/metabolism Cell Cycle Proteins Cell Line, Tumor Centrosome/metabolism Conserved Sequence Golgi Apparatus/metabolism Humans Intracellular Signaling Peptides and Proteins/chemistry,metabolism Mutant Proteins/metabolism Nerve Tissue Proteins/chemistry,metabolism Protein Binding Protein Transport Structure-Activity Relationship
Chemicals
CDK5RAP2 protein, human Calmodulin Cell Cycle Proteins Intracellular Signaling Peptides and Proteins Mutant Proteins Nerve Tissue Proteins
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Wang Zhe
Department of Biochemistry and State Key Laboratory of Molecular Neuroscience, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China.
Wu Tao
Shi Lin
Zhang Lin
Zheng Wei
Qu Jianan Y
Niu Ruifang
Qi Robert Z
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M110.105965
Published
2010-07-16
Epub
2010-00-13
Pages
22658-65
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2903348
Subset
IM
External Links
PubMed source DOI: 10.1074/jbc.M110.105965 PMC Full Text View journal details
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