Further definition of the sequence and position requirements of the arginine control element that mediates repression and induction by arginine in Saccharomyces cerevisiae.
Repression or induction of the genes involved in arginine biosynthesis or catabolism, respectively, both require participation of the ArgRp/Mcm1p regulatory complex. Our previous work showed that those opposite effects were mediated by a similar arginine-responsive element of 23 nucleotides (that we now call ARC, for ARginine Control) situated close to the start of transcription in the repressed promoters and far upstream of the TATA-element in the induced promoters. To define more precisely the sequence and position requirements of the ARC element, we have now characterized by mutagenesis the promoter elements of the arginine-repressible ARG1 and ARG8 genes. We also identify a functional ARC in the CPA1 promoter, thereby confirming, in agreement with our previous mRNA pulse-labelling data, the participation of a transcriptional component in the arginine regulation of that gene otherwise submitted to a translational regulation. From the 12 ARC elements now characterized, we have derived a consensus sequence and show that such a synthetic element is able to mediate ArgRp/Mcm1p-dependent arginine regulation. An important new finding illustrated by ARG1 and CPA1, is that contrary to what all the previous data suggested, repression can be mediated by ARC elements located far upstream of the TATA-box. The new data suggest that the arginine repressor might inhibit transcription in an active process.
Crabeel
M
M
Erfelijkheidsleer en Microbiologie, Vrije Universiteit Brussel and Onderzoekingsinstituut CERIA-COOVI, Belgium.
de Rijcke
M
M
Seneca
S
S
Heimberg
H
H
Pfeiffer
I
I
Matisova
A
A
D001120:Arginine / Q000378:metabolism*; D001124:Argininosuccinate Synthase / Q000378:metabolism; D001426:Bacterial Proteins; D001483:Base Sequence; D019733:Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor; D016384:Consensus Sequence / Q000235:genetics; D004268:DNA-Binding Proteins / Q000502:physiology; D005656:Fungal Proteins / Q000378:metabolism / Q000502:physiology; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D008025:Ligases / Q000378:metabolism; D026341:Minichromosome Maintenance 1 Protein; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011494:Protein Kinases / Q000502:physiology; D018085:Regulon / Q000235:genetics; D012097:Repressor Proteins / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA; D016385:TATA Box / Q000235:genetics; D015534:Trans-Activators / Q000502:physiology; D000637:Transaminases / Q000378:metabolism; D014157:Transcription Factors / Q000502:physiology; D014158:Transcription, Genetic / Q000235:genetics
C087018:ArgR protein, Bacteria; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D026341:Minichromosome Maintenance 1 Protein; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D001120:Arginine; D000637:Transaminases; C018731:acetylornithine transaminase; D011494:Protein Kinases; D008025:Ligases; D001124:Argininosuccinate Synthase; D019733:Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor; C064716:carbamoyl phosphate synthetase (arginine-specific)