In addition, microarray experiments revealed that high-level ureABC expression during nitrogen-limited growth was dependent upon PucR H. Jarmer and H. Saxild, unpublished observations. Nucleotide sequence of the ure P3 promoter.
The transcriptional start site is indicated by an asterisk. For unknown reasons, no induction was observed with allantoin data not shown. Taken together, these results indicate that ure P3 expression is activated by PucR.
This result indicates that TnrA is required for the activation of ure P3 promoter expression under these conditions. One site is centered 91 bp upstream of the ure P3 transcriptional start site, while the other site is centered 15 bp downstream of the transcriptional start site Fig.
The position of the upstream site is unusual in that the TnrA binding sites of other TnrA-activated promoters are centered 48 to 51 bp upstream of the transcriptional start site It has been shown previously that during nitrogen-limited growth the expression of pucR increases fold in a wild-type strain but only fourfold in a tnrA mutant 3.
As a result, nitrogen-limited tnrA mutants contain low but significant levels of PucR compared to wild-type cells. Taken together, these observations indicate that the defect in ure P3 expression observed in the tnrA mutant during nitrogen-limited growth results from its inability to synthesize wild-type levels of PucR. GlnR represses gene expression during growth on excess nitrogen sources such as glutamine The simplest model to explain these results is that GlnR binding to the downstream GlnR site inhibits the initiation of transcription at the ure P3 promoter and that the binding of GlnR to the downstream site can be strengthened by a cooperative interaction with GlnR bound to the upstream GlnR site.
By analogy to other systems 21 , GlnR-dependent regulation of ure P3 should be reduced when the distance between the upstream and downstream GlnR sites is changed by half-integral turns of DNA 5 bp. Moreover, deletion of a whole integral turn of DNA 10 bp should restore the correct phasing between these two sites and increase regulation by GlnR.
In contrast, expression of the URE66 fusion, which contains a full-integral-turn bp deletion between the two GlnR sites Fig.
The bp deletion in the URE66 fusion moves a sequence containing 7 consecutive A's closer to the ure P3 promoter. The higher levels of expression seen with the URE66 fusion may result from this sequence acting as an upstream promoter element that enhances transcription from the ure P3 promoter The regulation of the glnRA operon by GlnR has previously been shown to involve cooperative interactions.
Since GlnR binds simultaneously to both operators with in vitro and in vivo DNA footprinting experiments 5 , 11 , a cooperative interaction appears to occur between the GlnR dimers bound at these sites. This conclusion is supported by the observations that GlnR-dependent regulation of glnRA expression is completely relieved by either deletion of the upstream operator or insertion of a 5-bp DNA fragment between the two operators Although our analysis of the ure P3 promoter indicates that a cooperative interaction occurs between the forms of GlnR bound at the two GlnR sites in the ure P3 promoter region, GlnR still weakly regulates ure P3- lacZ fusions containing only the downstream GlnR site.
It is possible that GlnR binds with sufficient affinity to the downstream GlnR site in the ure P3 promoter to weakly regulate gene expression.
Alternatively, this low level of regulation may result from GlnR binding to both the downstream GlnR site and a yet unrecognized sequence with poor homology to a GlnR site in the ure P3 promoter region. Urease is widely distributed in bacteria The expression of urease is controlled by diverse regulatory mechanisms, suggesting that this enzyme has multiple physiological roles in bacteria.
For example, urease expression in Streptococcus salivarius is induced by growth at acidic pH 6. Since the degradation of urea to carbon dioxide and two molecules of ammonia produces a net increase in pH, high levels of urease can facilitate alkalization and bacterial survival in acidic environments. Because ammonia produced during urea catabolism can provide nitrogen for cell growth, the expression of urease in some bacteria is regulated in response to nitrogen availability.
During nitrogen-limited growth, urease expression is activated by the Ntr system in Klebsiella aerogenes Purine degradation occurs during nitrogen-limited growth in B. Since urea is produced by the final step in purine catabolism Fig.
Substrates for purine catabolism are likely obtained from the degradation of DNA present in the environment because B. It is also noteworthy that the expression of competence genes is induced during nitrogen-limited growth in B.
This observation raises the possibility that single DNA strands that are not incorporated into the chromosome during transformation may be degraded to provide nutrients for continued cell growth. Interestingly, the ability to utilize extracellular DNA as an energy source during stationary phase in Escherichia coli cells has been shown to be dependent upon homologs of proteins involved in transformation in Haemophilus influenzae and Neisseria gonorrhoeae 9.
Read article at publisher's site DOI : Kirst H , Kerfeld CA. Biochem Soc Trans , 49 3 , 01 Jun Cited by: 0 articles PMID: J Bacteriol , 19 , 08 Sep PLoS Genet , 14 9 :e, 28 Sep J Dairy Sci , 11 , 24 Aug Cited by: 4 articles PMID: To arrive at the top five similar articles we use a word-weighted algorithm to compare words from the Title and Abstract of each citation. J Bacteriol , 17 , 01 Sep J Bacteriol , 7 , 01 Apr Fisher SH.
Mol Microbiol , 32 2 , 01 Apr Cited by: articles PMID: J Bacteriol , 2 , 01 Jan Review Free to read. Contact us. Europe PMC requires Javascript to function effectively. Recent Activity. Search life-sciences literature Over 39 million articles, preprints and more Search Quick link: Coronavirus articles and preprints. Recent history Saved searches. Search articles by 'Jaclyn L Brandenburg'. Brandenburg JL 1 ,. Wray LV Jr ,.
Lars Beier Search articles by 'Lars Beier'. Beier L ,. Hanne Jarmer Search articles by 'Hanne Jarmer'. Jarmer H ,. Saxild HH ,. Affiliations 1 author 1. Share this article Share with email Share with twitter Share with linkedin Share with facebook. Abstract Expression of the P3 promoter of the Bacillus subtilis ureABC operon is activated during nitrogen-limited growth by PucR, the transcriptional regulator of the purine-degradative genes.
Free full text. J Bacteriol. PMID: Jaclyn L. Brandenburg , 1 Lewis V. Wray, Jr. Saxild , 2 and Susan H. Lewis V. Hans H. Susan H. Author information Article notes Copyright and License information Disclaimer. Phone: Fax: E-mail: ude. Received Jun 12; Accepted Aug 6. This article has been cited by other articles in PMC. Fax: E-mail: ude. Received May 9; Accepted Jul 5. All Rights Reserved.
This article has been cited by other articles in PMC. Go to:. Gel mobility shift assays. DNase I footprinting. Oligonucleotide mutagenesis. Enzyme assays. Table 1. CodY DNA binding affinities. Promoter Coeffector s a K 0. Open in a separate window. GTP was present only in the binding reaction mixture. Values are the averages from two or more independent experiments. The uncertainty is the standard error from nonlinear regression analysis of the data.
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