Nucleotide Sequence of the argR Gene of Escherichia coli K-12 and Isolation of Its Product, the Arginine Repressor

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 84; no. 19; pp. 6697 - 6701
• Main Authors: Lim, Dongbin, Oppenheim, Joel D., Eckhardt, Thomas, Maas, Werner K.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01.10.1987; National Acad Sciences
• Subjects: Amino Acid Sequence; Bacterial Proteins; Bacteriology; Base Sequence; beta-Galactosidase - genetics; beta-Galactosidase - isolation & purification; Biological and medical sciences; Enzymes; Escherichia coli - genetics; Escherichia coli Proteins; Fundamental and applied biological sciences. Psychology; Gels; Genes; Genes, Bacterial; Genes, Regulator; Genetics; Microbiology; Molecular Sequence Data; Molecular weight; Nucleotide sequences; Open reading frames; Plasmids; Promoter regions; Proteins; Reaction products; Recombinant Fusion Proteins - isolation & purification; Repressor Proteins - genetics; Repressor Proteins - isolation & purification; Sequencing; Transcription Factors - genetics; Gene product; Escherichia coli; Inheritance; Transcription repressor; Primary structure; Biosynthesis; Arginine; Gene; Aminoacid; Bacteria; Isolation; Escherichieae; Enterobacteriaceae
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.84.19.6697

In Escherichia coli, the arginine repressor, the product of the argR gene, in conjunction with L-arginine controls the synthesis of the enzymes of arginine biosynthesis. We describe the nucleotide sequence of the argR gene, including its control region, and show that formation of the repressor is autoregulated. The argR control region contains two promoters, one of which overlaps the operator site and, as with other arg genes, consists of two adjacent palindromic sequences (``ARG boxes''). The arginine repressor protein and an arginine repressor-β -galactosidase fusion protein were purified, and the amino acid sequence of the N-terminal end of the repressor protein portion of the fusion protein was determined. Antibodies prepared against the fusion protein react with the repressor. The repressor is precipitable by L-arginine, which facilitates its purification. The native repressor is a hexamer with a molecular weight of 98,000; its monomeric subunit has a molecular weight of 16,500. To verify its properties postulated from genetic studies, we show that in the presence of L-arginine, repressor inhibits transcription of argF and binds to the ARG boxes of argF and argR.