Determination of the DNA-binding sequences of ARGR proteins to arginine anabolic and catabolic promoters.
ARGRI, ARGRII, and ARGRIII proteins regulate the expression of arginine anabolic and catabolic genes. The integrity of these three proteins is required to observe the formation of a DNA-protein complex with the different promoters of arginine coregulated genes. A study of deletions and point mutations created in the 5' noncoding region of ARG3, ARG5,6, CAR1, and CAR2 genes shows that at least two regions, called BoxA and BoxB, are required for proper regulation of these genes by arginine and ARGR proteins. By gel retardation assay and DNase I footprinting analysis, we have determined precisely the target of the ARGR proteins. Sequences in and around BoxA are necessary for ARGR binding to these four promoters in vitro, whereas sequences in and around BoxB are clearly protected against DNase I digestion only for CAR1. Sequences present at BoxA and BoxB are well conserved among the four promoters. Moreover, pairing can occur between sequences at BoxA and BoxB which could lead to the creation of secondary structures in ARG3, ARG5,6, CAR1, and CAR2 promoters, favoring the binding of ARGR proteins in vivo.
Messenguy
F
F
Institut de Recherches du CERIA, Brussels, Belgium.
Dubois
E
E
Boonchird
C
C
D001120:Arginine / Q000378:metabolism*; D001483:Base Sequence; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics*; D003850:Deoxyribonuclease I; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D009710:Nucleotide Mapping; D015345:Oligonucleotide Probes; D017853:Phosphotransferases (Alcohol Group Acceptor); D010957:Plasmids; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins; D015183:Restriction Mapping; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors
C066955:ARG80 protein, S cerevisiae; C067875:ARG81 protein, S cerevisiae; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D015345:Oligonucleotide Probes; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D001120:Arginine; C425775:ARG82 protein, S cerevisiae; D017853:Phosphotransferases (Alcohol Group Acceptor); D003850:Deoxyribonuclease I