Nucleic acids encoding hBub1, a cell cycle checkpoint gene


Ontology type: sgo:Patent     


Patent Info

DATE

2002-01-01T00:00

AUTHORS

Wei Dai , Bin Ouyang , Huiqi Pan , Zhengdao Lan

ABSTRACT

Eukaryotic cells have evolved a mechanism that delays the onset of anaphase until chromosomes are properly positioned on the spindle. To understand the molecular basis of such surveillance mechanism in human cells, we have cloned a full-length cDNA encoding a putative mitotic checkpoint kinase termed hBub1. Sequence comparison reveals that hBub1 is a structurally conserved protein, sharing 23% amino acid residue identity with BUB1 of budding yeast. In addition, the N-terminal portion (161 amino acids) of hBub1 shows a significant homology to yeast MAD3, a protein also known to be involved in the mitotic checkpoint response pathway. Northern blot analyses show that hBub1 mRNA level is abundantly expressed in tissues or cells with a high mitotic index. When Dami cells undergoes terminal differentiation following treatment with phorbol ester, hBub1 expression in this cell line is rapidly downregulated. The hBub1 protein level is low in G1 and remains relatively constant in S, G2 and M phases. Immunofluorescence analysis shows that hBub1 protein co-localizes with a centromere-kinetochore antigen CREST in interphase, mitotic prophase and nocodazole-treated cells. Antibody electroporation experiments show that hBub1 is an important component of the spindle checkpoint pathway. Furthermore, fluorescence in situ hybridization analysis maps the hBub1 gene to chromosome 2q12-13. Our studies suggest that hBub1 expression is restricted to proliferating cells and appears to be involved in regulating cell-cycle progression. The molecular cloning of hBub1 cDNA will facilitate the study of its role in spindle checkpoint control as well as its potential role in certain genetic disorders. More... »

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