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Nucleic Acids Research, 1989, Vol. 17, No. 8 2947-2957
© 1989

MOLECULAR BIOLOGY

The narX and narL genes encoding the nitrate-sensing regulators of Escherichia coli are homologous to a family of prokaryotic two-component regulatory genes

Tsutomu Nohno, Sumihare Noji1, Shigehiko Taniguchi1 and Tauchi Saito

Department of Pharmacology, Kawasaki Medical School 577 Matsushima. Kurashiki 701-01, Japan 1Department of Biochemistry, Okayama University Dental School Shikata-cho 2-5-1, Okayama 700, Japan

Received February 2, 1989. Revised March 15, 1989. Accepted March 15, 1989.

The nucleotide sequence of a 4.4-kilobase SacII -SspI fragment encoding the narXL operon and a part of the narK gene of Escherichia coli has been determined. The narX and narL genes encode proteins of molecular weight 67,275 and 23,927, respectively, and are transcribed from a common promoter, narXp, locating within 429 bases upstream of narX. Transcription from narXp is not significantly induced by nitrate under anaerobiosis, whereas transcription from narK promoter, which overlaps narXp region and is transcribed divergently, is fully induced by nitrate. The N-terminal two-thirds of the NarL protein has extensive homology with those of a diverse set of prokaryotic regulatory proteins, including OmpR, PhoB, SfrA, UhpA, CheY, CheB, NtrC, DctD, FixJ, VirG, Spo0F, and Spo0A. A segment locating in the C-terminal half of the NarL protein seems to have potential most likely to form the helix-turn-helix structure characteristic of a class of DNA-binding protein. The protein is considered to play a role as a transcriptional activator of the nitrate reductase operon, narCHJI, and the narK gene. The C-terminal region of the NarX protein also has homology with other regulatory proteins known as counterparts of two-component regulatory systems, such as EnvZ, PhoR, PhoM, CpxA, NtrB, DctB, FixL, and VirA. Presence of two copies of hydrophobic segments in the N-terminal half of the NarX protein suggests the role as a transmembrane receptor sensing nitrate.


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