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

Journal Article · Thu May 01 04:00:00 EDT 2008 · Experimental Cell Research

DOI:https://doi.org/10.1016/j.yexcr.2008.02.021· OSTI ID:21128176

Yi, Wei; Enzhi, Shen; Na, Zhao; Qian, Liu; Jinling, Fan ^[1]; Marc, Jan ^[2]; Yongchao, Wang ^[1]; Le, Sun ^[3]

---

1. Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education, College of Life Sciences, Beijing Normal University, Beijing 100875 (China)
2. School of Biological Sciences, University of Sydney, Sydney 2006 (Australia)
3. Welson Pharmaceuticals, Haidian, Beijing 100085 (China)

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.

• OSTI ID: 21128176
• Journal Information: Experimental Cell Research, Journal Name: Experimental Cell Research Journal Issue: 8 Vol. 314; ISSN 0014-4827; ISSN ECREAL
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
AMINO ACID SEQUENCE
CELL CYCLE
CELL PROLIFERATION
CYTOPLASM
HELA CELLS
MASS SPECTROSCOPY
MICROSCOPY
MITOSIS
MONOCLONAL ANTIBODIES
PATIENTS
PROTEINS
RNA