Abstract
This paper investigates the effects of sense and antisense centromere/kinetochore complex protein-B (CENP-B) in cell cycle regulation. Full-length cenpb cDNA was subcloned into pBI-EGFP eukaryotic expression vector in both sense and antisense orientation. HeLa-Tet-Off cells were transfected with sense or antisense cenpb vectors. Sense transfection of HeLa-Tet-Off cells resulted in the formation of a large centromere/kinetochore complex, and apoptosis of cells following several times of cell division. A stable antisense cenpb transfected cell line, named HACPB, was obtained. The centromere/kinetochore complex of HACPB cells became smaller than control HeLa-Tet-Off cells and scattered, and the expression of CENP-B was down-regulated. In addition, delayed cell cycle progression, inhibited malignant phenotype, restrained ability of tumor formation in nude mice, and delayed entry from G2/M phase into next G1 phase were observed in HACPB cells. Furthermore, the expression of cyclin-dependent kinases (CDKs), cyclins, and CDK inhibitors (CKIs) were modulated during different phases of the cell cycle. CENP-B is an essential protein for the maintenance of the structure and function of centromere/kinetochore complex, and plays important roles in cell cycle regulation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Rattner, J. B., The structure of the mammalian centromere, BioEs- says, 1991, 13: 51–56.
Amon, A., The spindle checkpoint, Curr. Opin. Genet. Dev., 1999, 9: 69–75.
He, D., Zeng, C., Woods, K. et al., CENP-G: A new centromeric protein that is associated with the alpha-1 satellite DNA subfamily, Chromosoma, 1998, 107: 189 - 197.
Sugata, N., Li, S., Earnshaw, W. C. et al., Human CENP-H mul- timers colocalize with CENP-A and CENP-C at active centro- mere-kinetochore complexes, Hum. Mol. Genet., 2000, 9: 2919- 2926.
Nishihashi, A., Haraguchi, T., Hiraoka, Y. et al., CENP-I is essen- tial for centromere function in vertebrate cells, Dev. Cell, 2002, 2: 463–476.
Earnshaw, W. C., Sullivan, K. F., Machlin, P. S. et al., Molecular cloning of cDNA for CENP-B, the major human centromere autoantigen, J. Cell. Biol., 1987, 104: 817–829.
Kitagawa, K., Masumoto, H., Ikeda, M. et al., Analysis of pro- tein-DNA and protein-protein interactions of centromere protein B (CENP-B) and properties of the DNA-CENP-B complex in the cell cycle, Mol. Cell. Biol., 1995, 15: 1602–1612.
Ohzeki, J., Nakano, M., Okada, T. et al., CENP-B box is required for de novo centromere chromatin assembly on human alphoid DNA, J. Cell. Biol., 2002, 159: 765–775.
Zhang, H. X., Chen, D. G., Peng, A. et al., Relationship between CENP-B gene expression and the cell cycle, Acta Biologiae. Ex- perimentalis. Sinica, 1995, 28: 291–298.
Knehr, M., Poppe, M., Schroeter, D. et al., Cellular expression of human centromere protein C demonstrates a cyclic behavior with highest abundance in the G1 phase, Proc. Natl. Acad. Sci. USA, 1996, 93: 10234–10239.
Zeitlin, S. G., Barber, C. M., Allis, C. D. et al., Differential regula- tion of CENP-A and histone H3 phosphorylation in G2/M, J. Cell. Sci., 2001, 114:653–661.
Fukagawa, T., Mikami, Y., Nishihashi, A. et al., CENP-H, a con- stitutive centromere component, is required for centromere target- ing of CENP-C in vertebrate cells, EMBO. J., 2001, 20: 4603- 4617.
Weaver, B. A., Bonday, Z. Q., Putkey, F. R. et al., Centro- mere-associated protein-E is essential for the mammalian mitotic checkpoint to prevent aneuploidy due to single chromosome loss, J. Cell. Biol., 2003, 162: 551–563.
Liu, S. T., Hittle, J. C., Jablonski, S. A. et al., Human CENP-I specifies localization of CENP-F, MAD1 and MAD2 to kinetocho- res and is essential for mitosis, Nat. Cell. Biol., 2003, 5: 341–345.
Esguerra, R. L., Jia, L., Kaneko, T. et al., Immunohistochemical analysis of centromere protein F expression in buccal and gingival squamous cell carcinoma, Pathol. Int., 2004, 54: 82 - 89.
Liang, Q. J., Lu, X. F., Cheng, X. L. et al., The active expression of CenpB, a constitutive protein in the centromeres of chromosomes, in breast cancer tissues, Acta. Genetica. Sinica., 2004, 31: 236 - 240.
Liang, Q. J., Wang, Q., Lin, H. Y. et al., Effects of antisense CENP-B on the proliferation of HeLa (Tet-off) cells, Chinese Sci. Bul., 2002, 47(5): 379–383.
Vafa, O., Sullivan, K. F., Chromatin containing CENP-A and al- pha-satellite DNA is a major component of the inner kinetochore plate, Curr. Biol., 1997, 7: 897–900.
Fukagawa, T., Pendon, C., Morris, J. et al., CENP-C is necessary but not sufficient to induce formation of a functional centromere, EMBO. J., 1999, 18: 4196–4209.
Figueroa, J., Saffrich, R., Ansorge, W. et al., Microinjection of an- tibodies to centromere protein CENP-A arrests cells in interphase but does not prevent mitosis, Chromosoma., 1998, 107: 397–405.
Yen, T. J., Compton, D. A., Wise, D. et al., CENP-E, a novel hu- man centromere-associated protein required for progression from metaphase to anaphase, EMBO. J., 1991, 10: 1245–1254.
Author information
Authors and Affiliations
Corresponding author
Additional information
These two authors contributed equally to this work.
About this article
Cite this article
Luo, S., Lin, H., Qi, J. et al. Effects of sense and antisense centromere/kinetochore complex protein-B (CENP-B) in cell cycle regulation. Chin.Sci.Bull. 50, 2836–2843 (2005). https://doi.org/10.1360/982005-910
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1360/982005-910