PMID:8444804
| Citation |
Sun, D and Setlow, P (1993) Cloning, nucleotide sequence, and regulation of the Bacillus subtilis nadB gene and a nifS-like gene, both of which are essential for NAD biosynthesis. J. Bacteriol. 175:1423-32 |
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| Abstract |
A number of Bacillus subtilis genes involved in NAD biosynthesis have been cloned and sequenced. One of the genes encodes a polypeptide homologous to Escherichia coli L-aspartate oxidase, and its mutation resulted in a nicotinic acid (Nic)-dependent phenotype; this gene was termed nadB. A second open reading frame (orf2) was found downstream of nadB, and an insertional plasmid separating orf2 and nadB also gave a Nic-dependent phenotype. This result suggests that orf2 may also be involved in NAD biosynthesis and that nadB and orf2 are in the same operon. Upstream of nadB was a third gene, transcribed in the opposite direction to that of nadB-orf2. The amino acid sequence derived from the third gene was quite similar to those derived from nifS genes of various nitrogen-fixing bacteria; therefore, the third gene was termed nifS. As with nadB and orf2, mutations in nifS also resulted in a Nic-dependent phenotype. The promoter regions of nadB and nifS overlapped each other and both contained -10 and -35 sequences which resemble those of E sigma A-type promoters. Transcription from both the nifS and nadB promoters, as well as expression of a nadB-lacZ fusion, was repressed by Nic. However, nadB transcription and nadB-lacZ expression were decreased, at most, only slightly by a deletion in nifS. The possible role of the nifS gene product in NAD biosynthesis is discussed. |
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| Keywords |
Amino Acid Oxidoreductases/genetics; Amino Acid Sequence; Bacillus subtilis/genetics; Bacterial Proteins/genetics; Base Sequence; Cloning, Molecular; DNA, Bacterial; Gene Expression Regulation, Bacterial; Genes, Bacterial; Kinetics; Molecular Sequence Data; Mutation; NAD/biosynthesis; NAD/genetics; Nicotinamide Mononucleotide/analogs & derivatives; Nicotinamide Mononucleotide/biosynthesis; Nicotinamide Mononucleotide/genetics; Nitrogen Fixation/genetics; Restriction Mapping; Sequence Homology, Amino Acid |
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