Identification of a novel centrosomal protein Crp{sup F46} involved in cell cycle progression and mitosis

• Published in: Experimental cell research Vol. 314; no. 8
• Main Authors: Wei Yi, Shen Enzhi, Zhao Na, Liu Qian, Fan Jinling, Marc, Jan, Wang Yongchao, Sun Le, Liang Qianjin
• Format: Journal Article
• Language: English
• Published: United States 01.05.2008
• Subjects: 60 APPLIED LIFE SCIENCES; AMINO ACID SEQUENCE; CELL CYCLE; CELL PROLIFERATION; CYTOPLASM; HELA CELLS; MASS SPECTROSCOPY; MICROSCOPY; MITOSIS; MONOCLONAL ANTIBODIES; PATIENTS; PROTEINS; RNA
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/j.yexcr.2008.02.021

A novel centrosome-related protein Crp{sup F46} was detected using a serum F46 from a patient suffering from progressive systemic sclerosis. We identified the protein by immunoprecipitation and Western blotting followed by tandem mass spectrometry sequencing. The protein Crp{sup F46} has an apparent molecular mass of {approx} 60 kDa, is highly homologous to a 527 amino acid sequence of the C-terminal portion of the protein Golgin-245, and appears to be a splice variant of Golgin-245. Immunofluorescence microscopy of synchronized HeLa cells labeled with an anti-Crp{sup F46} monoclonal antibody revealed that Crp{sup F46} localized exclusively to the centrosome during interphase, although it dispersed throughout the cytoplasm at the onset of mitosis. Domain analysis using Crp{sup F46} fragments in GFP-expression vectors transformed into HeLa cells revealed that centrosomal targeting is conferred by a C-terminal coiled-coil domain. Antisense Crp{sup F46} knockdown inhibited cell growth and proliferation and the cell cycle typically stalled at S phase. The knockdown also resulted in the formation of poly-centrosomal and multinucleate cells, which finally became apoptotic. These results suggest that Crp{sup F46} is a novel centrosome-related protein that associates with the centrosome in a cell cycle-dependent manner and is involved in the progression of the cell cycle and M phase mechanism.