Distinct and redundant roles of the two protein kinase A isoforms Tpk1p and Tpk2p in morphogenesis and growth of Candida albicans.
TPK1 and TPK2 encode both isoforms of protein kinase A (PKA) catalytic subunits in Candida albicans. Mutants lacking both TPK1 alleles showed defective hyphal morphogenesis on solid inducing media, whereas in liquid hypha, formation was affected slightly. In contrast, tpk2 mutants were only partially morphogenesis defective on solid media, whereas a strong block was observed in liquid. In addition, the yeast forms of tpk2-- but not tpk1-- mutants were completely deficient in invading agar. Because Tpk1p and Tpk2p differ in their N-terminal domains of approximately 80--90 amino acids, while the catalytic portions are highly homologous, the functions of hybrid Tpk proteins with exchanged N-terminal domains were tested. The results demonstrate that the catalytic portions mediate Tpk protein specificities with regard to filamentation, whereas agar invasion is mediated by the N-terminal domain of Tpk2p. Homozygous tpk1 and tpk2 mutants grew normally; however, a tpk2 mutant strain containing a single regulatable TPK1 allele (PCK1p-TPK1) at low expression levels was severely growth defective. It was completely blocked in hyphal morphogenesis and was stress resistant to high osmolarities or temperatures. Thus, both Tpk isoforms in C. albicans share growth functions but, unlike Saccharomyces cerevisiae isoforms, they have positive, specific roles in filament formation in different environments.
Bockmühl
D P
DP
Institut für Mikrobiologie, Heinrich-Heine-Universität, Universitätsstr. 1/26.12, 40225 Düsseldorf, Germany.
Krishnamurthy
S
S
Gerads
M
M
Sonneborn
A
A
Ernst
J F
JF
D000483:Alleles; D002176:Candida albicans / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics*; D017931:DNA Primers; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D007527:Isoenzymes / Q000235:genetics; D007700:Kinetics; D009024:Morphogenesis; D010455:Peptides; D016133:Polymerase Chain Reaction; D021122:Protein Subunits; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D007527:Isoenzymes; D010455:Peptides; D021122:Protein Subunits; C097188:polyglutamine; D017868:Cyclic AMP-Dependent Protein Kinases