Evidence for involvement of Saccharomyces cerevisiae protein kinase C in glucose induction of HXT genes and derepression of SUC2.
The PKC1 gene in the yeast Saccharomyces cerevisiae encodes protein kinase C that is known to control a mitogen-activated protein (MAP) kinase cascade consisting of Bck1, Mkk1 and Mkk2, and Mpk1. This cascade affects the cell wall integrity but the phenotype of Pkc1 mutants suggests additional targets which have not yet been identified. We show that a pkc1Delta mutant, as opposed to mutants in the MAP kinase cascade, displays two major defects in the control of carbon metabolism. It shows a delay in the initiation of fermentation upon addition of glucose and a defect in derepression of SUC2 gene after exhaustion of glucose from the medium. After addition of glucose the production of both ethanol and glycerol started very slowly. The V(max) of glucose transport dropped considerably and Northern blot analysis showed that induction of the HXT1, HXT2 and HXT4 genes was strongly reduced. Growth of the pkc1Delta mutant was absent on glycerol and poor on galactose and raffinose. Oxygen uptake was barely present. Derepression of invertase activity and SUC2 transcription upon transfer of cells from glucose to raffinose was deficient in the pkc1Delta mutant as opposed to the wild-type. Our results suggest an involvement of Pkc1p in the control of carbon metabolism which is not shared by the downstream MAP kinase cascade.
Brandão
R L
RL
Laboratório de Bioquímica Celular e Molecular, Núcleo de Pesquisas em Ciências Biológicas, Escola de Farmácia, Universidade Federal de Ouro Preto, Campus do Morro do Cruzeiro, Brazil. rlbrand@nupeb.ufop.br
Etchebehere
L
L
Queiroz
C C
CC
Trópia
M J
MJ
Ernandes
J R
JR
Gonçalves
T
T
Loureiro-Dias
M C
MC
Winderickx
J
J
Thevelein
J M
JM
Leiper
F C
FC
Carling
D
D
Castro
I M
IM
D051792:Basic Helix-Loop-Helix Proteins; D015152:Blotting, Northern; D015966:Gene Expression Regulation, Fungal; D005947:Glucose / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000235:genetics*; D009154:Mutation; D011493:Protein Kinase C / Q000235:genetics / Q000502:physiology*; D012097:Repressor Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D015398:Signal Transduction / Q000502:physiology; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics; D014158:Transcription, Genetic; D043324:beta-Fructofuranosidase
D051792:Basic Helix-Loop-Helix Proteins; D005947:Glucose; D006026:Glycoside Hydrolases; D011493:Protein Kinase C; D012097:Repressor Proteins; D014157:Transcription Factors; D043324:beta-Fructofuranosidase; C095572:Hand1 protein, mouse