Mutants affected in the putative diacylglycerol binding site of yeast protein kinase C.
In an attempt to study the functional similarities between protein kinase C from the yeast Saccharomyces cerevisiae and its human homologues we have started in vitro mutagenesis to alter specific domains. Here we report on the exchange of four cysteine residues by serines in yeast Pkc1p that have been shown to be essential for diacylglycerol (DAG) binding and activation by this compound in humans. The mutant yeast protein leads to sensitivity to caffeine and low concentrations of SDS when expressed in a pkc1 deletion strain. However, sensitivity to staurosporine was not affected. Our data indicate that the conserved DAG binding domain serves an important function in yeast Pkc1p.
Jacoby
J J
JJ
Institut für Mikrobiologie, Heinrich-Heine-Universität Düsseldorf, Germany.
Schmitz
H P
HP
Heinisch
J J
JJ
D000595:Amino Acid Sequence; D001557:Benzenesulfonates / Q000494:pharmacology; D001665:Binding Sites; D002110:Caffeine / Q000494:pharmacology; D004075:Diglycerides / Q000378:metabolism*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011493:Protein Kinase C / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D019311:Staurosporine / Q000494:pharmacology; D013329:Structure-Activity Relationship
D001557:Benzenesulfonates; D004075:Diglycerides; D012967:Sodium Dodecyl Sulfate; D002110:Caffeine; C007061:C.I. Fluorescent Brightening Agent 28; D011493:Protein Kinase C; D019311:Staurosporine