Physiological and transcriptional characterization of Saccharomyces cerevisiae strains with modified expression of catabolic regulators.
A comparative physiological and transcriptional study is presented on wild-type Saccharomyces cerevisiae and mutants with altered levels of catabolic regulators: hxk2Delta lacking hexokinase2, HAP4 / overproducing hap4p and hxk2 Delta HAP4 upward arrow. Relative to the wild-type, HAP4 / showed the same growth rate with some increased yield on glucose, and hxk2Delta grew 28% slower but with a dramatically improved yield. Hxk2 Delta HAP4 / grew 14% slower but showed fully oxidative growth. A higher yield correlated with increased respiration. For both hxk2 Delta strains, glucose repression was suppressed (upregulation of high-affinity sugar transporters, invertase and oxidative phosphorylation). T-profiler analysis showed that genes under control of the hap2/3/4/5-binding motif were significantly altered in expression in all strains. HAP4 overexpression, directly or in hxk2 knockouts, led to repression of the genes containing the Zap1p motif including ZAP1 itself, indicating altered zinc metabolism. Whereas HAP4 overexpression resulted in a shift towards oxidative metabolism only, deletion of HXK2 resulted in a strain that, in addition to being oxidative, almost completely lacked the ability to sense glucose. As the double mutant had an energy efficiency close to the maximum even with excess glucose and was derepressed to a larger extent and over a broader range, the functioning of the two regulators is in general considered to be additive.
Schuurmans
J Merijn
JM
Swammerdam Institute for Life Sciences, BioCentrum Amsterdam, Faculty of Science, University of Amsterdam, Amsterdam, The Netherlands.
Boorsma
André
A
Lascaris
Romeo
R
Hellingwerf
Klaas J
KJ
Teixeira de Mattos
M Joost
MJ
D023081:CCAAT-Binding Factor / Q000378:metabolism; D004789:Enzyme Activation; D015966:Gene Expression Regulation, Fungal; D005809:Genes, Regulator; D006593:Hexokinase / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic
D023081:CCAAT-Binding Factor; C061437:HAP4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C490214:HXK2 protein, S cerevisiae; D006593:Hexokinase