Transcriptional control of the glnD gene is not dependent on nitrogen availability in Escherichia coli.
Glutamine synthetase (GS) is one of the most important enzymes in the assimilation of nitrogenous compounds in Escherichia coli and related bacteria. For the control of its activity and biosynthesis, tricyclic cascades of uridylylation/deuridylylation of PII protein, adenylylation/deadenylylation of glutamine synthetase, and phosphorylation/dephosphorylation of Ntr1 are operating, where the regulation of uridylylation/deuridylylation by uridylyl transferase-uridylyl removing enzyme (UT-UR) (the product of the glnD gene) would play the ultimate nitrogen sensing role. However, the possible nitrogen-regulatable element in the upstream of the glnD gene has been debated. In the present experiment, we have cloned and sequenced the four minute regions of the Escherichia coli chromosome, where rpsB, map, glnD, and dapD genes have been identified in sequence. We could localize the transcriptional start site at seven nucleotides upstream of the translation initiation codon by primer extension analysis. The nitrogen dependency of the glnD gene has been analyzed by Northern blot, RNase protection, and promoter-luciferase activity assays. These data suggested a constitutive expression of the glnD gene independent of nitrogen availability. From these results, it could be concluded that the ultimate nitrogen sensing device for the bacteria should be the UT-UR itself, through modulation of its activity in response to the nitrogen status rather than its biosynthetic mechanism.
Kim
I H
IH
Department of Biochemistry, Dong-A University College of Medicine, Pusan, Korea.
Kwak
S J
SJ
Kang
J
J
Park
S C
SC
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism* / Q000494:pharmacology; D009713:Nucleotidyltransferases / Q000235:genetics* / Q000378:metabolism; D051922:PII Nitrogen Regulatory Proteins; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics
D004269:DNA, Bacterial; D051922:PII Nitrogen Regulatory Proteins; D012333:RNA, Messenger; D009713:Nucleotidyltransferases; C022515:regulatory protein uridylyltransferase; D009584:Nitrogen