Generation of a set of conditional analog-sensitive alleles of essential protein kinases in the fission yeast Schizosaccharomyces pombe.
The genome of the fission yeast Schizosaccharomyces pombe encodes for 17 protein kinases that are essential for viability. Studies of the essential kinases often require the use of mutant strains carrying conditional alleles. To inactivate these kinases conditionally, we applied a recently developed chemical genetic strategy. The mutation of a single residue in the ATP-binding pocket confers sensitivity to small-molecule inhibitors, allowing for specific inactivation of the modified kinase. Using this approach, we constructed conditional analog-sensitive alleles of 13 essential protein kinases in the fission yeast S. pombe.
Max F Perutz Laboratories, University of Vienna, Vienna, Austria.
Lubos
L
Cipak
Chao
C
Zhang
Ines
I
Kovacikova
Cornelia
C
Rumpf
Eva
E
Miadokova
Kevan M
KM
Shokat
Juraj
J
Gregan
D000255:Adenosine Triphosphate / Q000378:metabolism; D000483:Alleles; D001665:Binding Sites / Q000235:genetics; D002934:Cinnamates; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli; D055786:Gene Knockout Techniques; D020043:Genes, Essential / Q000235:genetics; D006921:Hygromycin B / Q000031:analogs & derivatives; D008954:Models, Biological; D015394:Molecular Structure; D009154:Mutation / Q000235:genetics; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism; D012568:Schizosaccharomyces / Q000201:enzymology*; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D013309:Streptothricins
D002934:Cinnamates; D017931:DNA Primers; D006921:Hygromycin B; C026273:hygromycin A; D013309:Streptothricins; D000255:Adenosine Triphosphate; D011494:Protein Kinases