title: Conserved motif of CDK5RAP2 mediates its localization to centrosomes and the Golgi complex.
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.
authors.affiliation: Department of Biochemistry and State Key Laboratory of Molecular Neuroscience, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Wang
authors.forename: Tao
authors.initials: T
authors.lastname: Wu
authors.forename: Lin
authors.initials: L
authors.lastname: Shi
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Zheng
authors.forename: Jianan Y
authors.initials: JY
authors.lastname: Qu
authors.forename: Ruifang
authors.initials: R
authors.lastname: Niu
authors.forename: Robert Z
authors.initials: RZ
authors.lastname: Qi
mesh_terms: D020816:Amino Acid Motifs; D002147:Calmodulin / Q000378:metabolism; D018797:Cell Cycle Proteins; D045744:Cell Line, Tumor; D018385:Centrosome / Q000378:metabolism*; D017124:Conserved Sequence; D006056:Golgi Apparatus / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000378:metabolism*; D050505:Mutant Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D013329:Structure-Activity Relationship
chemical_list: C405929:CDK5RAP2 protein, human; D002147:Calmodulin; D018797:Cell Cycle Proteins; D047908:Intracellular Signaling Peptides and Proteins; D050505:Mutant Proteins; D009419:Nerve Tissue Proteins
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