Coordinate synthesis of the enzymes of pyrimidine biosynthesis in Bacillus subtilis.

Paulus, Thomas J.; McGarry, Thomas J.; Shekelle, Paul G; Rosenzweig, Simon; Switzer, Robert L.

doi:10.1128/jb.149.2.775-778.1982

Cited by 23 publications

(7 citation statements)

References 11 publications

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“…ratio 0.09 ± 0.04) in the presence of C-6-H compared with the control. This whole operon is regulated by pyrR and strongly depends on the presence of pyrimidine nucleotides (Turner et al 1994 ; Paulus et al 1982 ). Synthesis of pyrimidine is crucial for the cells to divide and therefore cell survival (Turnbough and Switzer 2008 ).…”

Section: Resultsmentioning

confidence: 99%

The effect of skin fatty acids on Staphylococcus aureus

et al. 2014

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Staphylococcus aureus is a commensal of the human nose and skin. Human skin fatty acids, in particular cis-6-hexadecenoic acid (C-6-H), have high antistaphylococcal activity and can inhibit virulence determinant production. Here, we show that sub-MIC levels of C-6-H result in induction of increased resistance. The mechanism(s) of C-6-H activity was investigated by combined transcriptome and proteome analyses. Proteome analysis demonstrated a pleiotropic effect of C-6-H on virulence determinant production. In response to C-6-H, transcriptomics revealed altered expression of over 500 genes, involved in many aspects of virulence and cellular physiology. The expression of toxins (hla, hlb,hlgBC) was reduced, whereas that of host defence evasion components (cap, sspAB, katA) was increased. In particular, members of the SaeRS regulon had highly reduced expression, and the use of specific mutants revealed that the effect on toxin production is likely mediated via SaeRS. Electronic supplementary materialThe online version of this article (doi:10.1007/s00203-014-1048-1) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

The effect of skin fatty acids on Staphylococcus aureus

et al. 2014

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“…The genetically unlinked pyr genes of E. coli and S. typhimurium are noncoordinately regulated by various mechanisms, including transcriptional and translational attenuation (11,31,48,52). In B. subtilis, the pyrimidine biosynthesis genes are clustered and their expression is regulated coordinately by pyrimidines (36,47). We have previously shown that expression of pyrB, pyrE, and pyrF in B. caldolyticus is repressed seven-to ninefold by the addition of uracil to the growth medium (13).…”

Section: Discussionmentioning

confidence: 99%

The pyrimidine biosynthesis operon of the thermophile Bacillus caldolyticus includes genes for uracil phosphoribosyltransferase and uracil permease

Ghim

Neuhard

1994

J Bacteriol

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A 3-kb DNA segment of the Bacillus caldolyticus genome including the 5' end end of the pyr cluster has been cloned and sequenced. The sequence revealed the presence of two open reading frames, pyrR and pyrP, located immediately upstream of the previously sequenced pyrB gene encoding the pyrimidine biosynthesis enzyme aspartate transcarbamoylase. The pyrR and pyrP genes encoded polypeptides with calculated molecular masses of 19.9 and 45.2 kDa, respectively. Expression of these ORFs was confirmed by analysis of plasmid-encoded polypeptides in minicells. Sequence alignment and complementation analyses identified the pyrR gene product as a uracil phosphoribosyltransferase and the pyrP gene product as a membrane-bound uracil permease. By using promoter expression vectors, a 650-bp EcoRI-HincII fragment, including the 5' end of pyrR and its upstream region, was found to contain the pyr operon promoter. The transcriptional start point was located by primer extension at a position 153 bp upstream of the pyrR translation initiation codon, 7 bp 3' of a sequence resembling a sigma A-dependent Bacillus subtilis promoter. This established the following organization of the ten cistrons within the pyr operon: promoter-pyrR-pyrP-pyrB-pyrC-pyrAa-pyrA b-orf2-pyrD-pyrF-pyrE. The nucleotide sequences of the region upstream of pyrR and of the pyrR-pyrP and pyrP-pyrB intercistronic regions indicated that the transcript may form two mutually exclusive secondary structures within each of these regions. One of these structures resembled a rho-independent transcriptional terminator. The possible implication of these structures for pyrimidine regulation of the operon is discussed.

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“…The nucleotide sequence of the entire B. subtilis pyr gene cluster has been determined previously (24, 37). All of the genes in the cluster appear to be expressed as a single transcriptional unit; pyr gene expression is coordinate and is strongly regulated by pyrimidines (33). Two putative transcription terminators, similar in structure to E. coli rho-independent terminators, were identified in the 5' end of the pyr cluster, one immediately downstream from the point of transcription initiation (37) and the other immediately preceding the pyrB gene (24).…”

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confidence: 99%

Regulation of the Bacillus subtilis pyrimidine biosynthetic (pyr) gene cluster by an autogenous transcriptional attenuation mechanism

1994

Self Cite

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A complete transcript of the Bacillus subtilis pyr operon contains the following elements in 5' to 3' order: a 151-nucleotide (nt) untranslated leader; pyrR, encoding a 20-kDa protein; a 173-nt intercistronic region; pyrP, encoding a 46-kDa protein; a 145-nt intercistronic region; and eight overlapping cistrons encoding all of the six enzymes for de novo pyrimidine biosynthesis. Transcription is controlled by the availability of pyrimidines via an attenuation mechanism. There are three transcription terminators within the operon, each of which is preceded by another stem-loop structure, the antiterminator, whose formation would prevent formation of the terminator stem-loop. These are located in the leader, the pyrR-pyrP intercistronic region, and the pyrP-pyrB intercistronic region. Northern (RNA) blot analysis has identified transcripts of lengths which coincide with termination at these proposed attenuation sites and whose relative abundances vary in the expected pyrimidine-dependent manner. Each antiterminator contains a 50-base conserved sequence in its promoter-proximal half. Various transcriptional fusions of the pyr promoter and surrounding sequences to promoterless reporter genes support an attenuation mechanism whereby when pyrimidines are abundant, the PyrR protein binds to the conserved sequence in the pyr mRNA and disrupts the antiterminator, permitting terminator hairpin formation and promoting transcription termination. Deletion of pyrR from the chromosome resulted in the constitutive, elevated expression of aspartate transcarbamylase, which is encoded by pyrB, the third gene in the operon. Complementation of an E. coli upp mutant, as well as direct enzymatic assay, has demonstrated that pyrR also confers uracil phosphoribosyltransferase activity. Analysis of pyrR and upp deletion mutants demonstrated that upp, not pyrR, encodes the quantitatively important uracil phosphoribosyltransferase activity. The pyrP gene probably encodes an integral membrane uracil permease.

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Coordinate synthesis of the enzymes of pyrimidine biosynthesis in Bacillus subtilis.

Cited by 23 publications

References 11 publications

The effect of skin fatty acids on Staphylococcus aureus

The effect of skin fatty acids on Staphylococcus aureus

The pyrimidine biosynthesis operon of the thermophile Bacillus caldolyticus includes genes for uracil phosphoribosyltransferase and uracil permease

Regulation of the Bacillus subtilis pyrimidine biosynthetic (pyr) gene cluster by an autogenous transcriptional attenuation mechanism

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