Creation and characterization of temperature-sensitive CENP-C mutants in vertebrate cells
AUTOR(ES)
Fukagawa, Tatsuo
FONTE
Oxford University Press
RESUMO
CENP-C is an evolutionarily conserved centromere protein that is thought to be an important component in kinetochore assembly in vertebrate cells. However, the functional role of CENP-C in cell cycle progression remains unclear. To further understand CENP-C function, we developed a method incorporating the hyper-recombinogenic chicken B lymphocyte cell line DT40 to create several temperature-sensitive CENP-C mutants in DT40 cells. We found that, under restrictive conditions, one temperature-sensitive mutant, ts4-11, displayed metaphase delay and chromosome missegregation but proceeded through the cell cycle until arrest at G1 phase. Furthermore, ts4-11 cells were transfected with a human HeLa cell cDNA library maintained in a retroviral vector, and genes that suppressed the temperature-sensitive phenotype were identified. One of these suppressor genes encodes SUMO-1, which is a ubiquitin-like protein. This finding suggests that SUMO-1 may be involved in centromere function in vertebrate cells. The novel strategy reported here will be useful and applicable to a wide range of proteins that have general cell-autonomous function in vertebrate cells.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=55920Documentos Relacionados
- CENP-H, a constitutive centromere component, is required for centromere targeting of CENP-C in vertebrate cells
- Creation and characterization of temperature-sensitive mutants of the lck tyrosine protein kinase.
- Isolation and Preliminary Characterization of Temperature-Sensitive Mutants of Rhizobiophage C
- Domains required for CENP-C assembly at the kinetochore.
- Isolation and Characterization of Sendai Virus Temperature-Sensitive Mutants
