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Research article

Cep57, a multidomain protein with unique microtubule and centrosomal localization domains

Ko Momotani, Alexander S. Khromov, Tsuyoshi Miyake, P. Todd Stukenberg, Avril V. Somlyo
Biochemical Journal Jun 01, 2008, 412 (2) 265-273; DOI: 10.1042/BJ20071501
Ko Momotani
Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA 22908, U.S.A.
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Alexander S. Khromov
Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA 22908, U.S.A.
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Tsuyoshi Miyake
Department of Microbiology, University of Virginia, Charlottesville, VA 22908, U.S.A.
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P. Todd Stukenberg
Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, VA 22908, U.S.A.
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Avril V. Somlyo
Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA 22908, U.S.A.
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  • For correspondence: avs5u@virginia.edu
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Abstract

The present study demonstrates different functional domains of a recently described centrosomal protein, Cep57 (centrosomal protein 57). Endogenous Cep57 protein and ectopic expression of full-length protein or the N-terminal coiled-coil domain localize to the centrosome internal to γ-tubulin, suggesting that it is either on both centrioles or on a centromatrix component. The N-terminus can also multimerize with the N-terminus of other Cep57 molecules. The C-terminus contains a second coiled-coil domain that directly binds to MTs (microtubules). This domain both nucleates and bundles MTs in vitro. This activity was also seen in vivo, as overexpression of full-length Cep57 or the C-terminus generates nocodazole-resistant MT cables in cells. Based on the present findings, we propose that Cep57 serves as a link with its N-terminus anchored to the centriole or centromatrix and its C-terminus to MTs.

  • centriole
  • centromatrix
  • centrosomal protein 57 (Cep57)
  • centrosome
  • electron microscopy
  • microtubule formation nucleation

Abbreviations: AKAP450, A-kinase-anchoring protein of 450 kDa; Cep57, centrosomal protein 57; Cep57R, Cep57-related protein; CLD, centrosome localization domain; CNN, centrosomin; Cy3, indocarbocyanine; DMEM, Dulbecco's modified Eagle's medium; DsRed, Discosoma corallimorpharian red fluorescent protein; EGF, epidermal growth factor; EM, electron microscopy; FBS, fetal bovine serum; GFP, green fluorescent protein; EGFP, enhanced GFP; GST, glutathione transferase; HEK-293 cells, human embryonic kidney-293 cells; hUbC, human ubiquitin C; JRed, a red fluorescent protein from a jellyfish of the suborder Anthomedusae; MT, microtubule; RNAi, RNA interference; SPD-5, spindle-defective protein-5

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June 2008

Volume: 412 Issue: 2

Biochemical Journal: 412 (2)
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Cep57, a multidomain protein with unique microtubule and centrosomal localization domains
Ko Momotani, Alexander S. Khromov, Tsuyoshi Miyake, P. Todd Stukenberg, Avril V. Somlyo
Biochemical Journal Jun 2008, 412 (2) 265-273; DOI: 10.1042/BJ20071501
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Cep57, a multidomain protein with unique microtubule and centrosomal localization domains
Ko Momotani, Alexander S. Khromov, Tsuyoshi Miyake, P. Todd Stukenberg, Avril V. Somlyo
Biochemical Journal Jun 2008, 412 (2) 265-273; DOI: 10.1042/BJ20071501

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Keywords

centriole
centromatrix
centrosomal protein 57 (Cep57)
centrosome
electron microscopy
microtubule formation nucleation

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