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Expression of a dominant negative allele of cdc2 prevents activation of the endogenous p34cdc2 kinase (1991)

Fleig, Ursula N. ; Nurse, Paul

Springer

Molecular genetics and genomics 226 (1991), S. 432-440

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ISSN: 1617-4623

Keywords: Schizosaccharomyces pombe ; cdc2 protein kinase ; Dominant negative allele

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Summary The cdc2 gene of the fission yeast Schizosaccharomyces pombe encodes a 34 kDa phosphoprotein with serine/threonine protein kinase activity that acts as the key component in regulation of the eukaryotic cell cycle. We used a repressible promoter fused to the cdc2 cDNA to isolate conditionally dominant negative mutants of cdc2. One of these mutants, DL5, is described in this paper. Overexpression of the mutant protein in a wild-type cdc2 background is lethal and confers cell cycle arrest with a typical cdc phenotype. Sequencing of the mutant cdc2 gene revealed a single amino acid substitution in a region highly conserved in cdc2-like proteins. The mutant protein exhibits no protein kinase activity, but is able to bind a component(s) required for an active protein kinase complex and thereby prevents binding of this component(s) to the co-existing wild-type cdc2 protein. We also demonstrate that S. pombe p34cdc2 contains no phosphoserine.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00260656

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The centromere of budding yeast (1993)

Hegemann, Johannes H. ; Fleig, Ursula N.

New York, NY : Wiley-Blackwell

BioEssays 15 (1993), S. 451-460

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ISSN: 0265-9247

Keywords: Life and Medical Sciences ; Cell & Developmental Biology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology , Medicine

Notes: Stable maintenance of genetic information during meiosis and mitosis is dependent on accurate chromosome transmission. The centromere is a key component of the segregational machinery that couples chromosomes with the spindle apparatus. Most of what is known about the structure and function of the centromeres has been derived from studies on yeast cells. In Saccharomyces cerevisiae, the centromere DNA requirements for mitotic centromere function have been defined and some of the proteins required for an active complex have been identified. Centromere DNA and the centromere proteins form a complex that has been studied extensively at the chromatin level. Finally, recent findings suggest that assembly and activation of the centromere are integrated in tethe cell cycle.

Additional Material: 6 Ill.