Analysis of Bacillus subtilis tag gene expression using transcriptional fusions

Mauël, Catherine; Young, Michael; Monsutti-Grecescu, Alice; Marriott, Shirley A.; Karamata, Dimitri

doi:10.1099/13500872-140-9-2279

Cited by 36 publications

(18 citation statements)

References 45 publications

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“…However, in contrast to Lang et al and in agreement with Soldo et al (40) and Lahooti and Harwood (25), we found that phosphate comprises a significant fraction of the anionic component in the walls of phosphate-limited cells. Our values are consistent with observations that the level of tag gene expression is one-third or one-half of the normal level during phosphate limitation (28,32). Together, these data support a model in which substantial quantities of teichoic acid continue to be incorporated into the wall.…”

Section: Discussionsupporting

confidence: 80%

“…Teichuronic acid is a phosphate-free carbohydrate polymer containing glucuronic acid (43) and is synthesized by enzymes encoded in the tua operon (40). When phosphate is limited, the PhoP/PhoR system induces transcription of the tua operon (29) and represses transcription of the tag divergon (28,32). Grant demonstrated that B. subtilis releases its existing teichoic acid into the medium and takes in phosphate from the medium (16).…”

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Teichoic Acid Is an Essential Polymer in Bacillus subtilis That Is Functionally Distinct from Teichuronic Acid

et al. 2004

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Wall teichoic acids are anionic, phosphate-rich polymers linked to the peptidoglycan of gram-positive bacteria. In Bacillus subtilis, the predominant wall teichoic acid types are poly(glycerol phosphate) in strain 168 and poly(ribitol phosphate) in strain W23, and they are synthesized by the tag and tar gene products, respectively. Growing evidence suggests that wall teichoic acids are essential in B. subtilis; however, it is widely believed that teichoic acids are dispensable under phosphate-limiting conditions. In the work reported here, we carefully studied the dispensability of teichoic acid under phosphate-limiting conditions by constructing three new mutants. These strains, having precise deletions in tagB, tagF, and tarD, were dependent on xyloseinducible complementation from a distal locus (amyE) for growth. The tarD deletion interrupted poly(ribitol phosphate) synthesis in B. subtilis and represents a unique deletion of a tar gene. When teichoic acid biosynthetic proteins were depleted, the mutants showed a coccoid morphology and cell wall thickening. The new wall teichoic acid biogenesis mutants generated in this work and a previously reported tagD mutant were not viable under phosphate-limiting conditions in the absence of complementation. Cell wall analysis of B. subtilis grown under phosphate-limited conditions showed that teichoic acid contributed approximately one-third of the wall anionic content. These data suggest that wall teichoic acid has an essential function in B. subtilis that cannot be replaced by teichuronic acid.

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confidence: 80%

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Teichoic Acid Is an Essential Polymer in Bacillus subtilis That Is Functionally Distinct from Teichuronic Acid

et al. 2004

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“…significantly down-regulated in response to phosphate starvation (44). However, several, but not all of the genes in the tua operon were included in the outlier analysis as being induced in response to phosphate starvation.…”

Section: Resultsmentioning

confidence: 99%

Genome-Wide Transcriptional Analysis of the Phosphate Starvation Stimulon ofBacillus subtilis

Allenby¹,

O'Connor²,

Prágai³

et al. 2005

J Bacteriol

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Bacillus subtilis responds to phosphate starvation stress by inducing the PhoP and SigB regulons. While the PhoP regulon provides a specific response to phosphate starvation stress, maximizing the acquisition of phosphate (P i ) from the environment and reducing the cellular requirement for this essential nutrient, the SigB regulon provides nonspecific resistance to stress by protecting essential cellular components, such as DNA and membranes. We have characterized the phosphate starvation stress response of B. subtilis at a genome-wide level using DNA macroarrays. A combination of outlier and cluster analyses identified putative new members of the PhoP regulon, namely, yfkN (2,3 cyclic nucleotide 2-phosphodiesterase), yurI (RNase), yjdB (unknown), and vpr (extracellular serine protease). YurI is thought to be responsible for the nonspecific degradation of RNA, while the activity of YfkN on various nucleotide phosphates suggests that it could act on substrates liberated by YurI, which produces 3 or 5 phosphoribonucleotides. The putative new PhoP regulon members are either known or predicted to be secreted and are likely to be important for the recovery of inorganic phosphate from a variety of organic sources of phosphate in the environment.When Bacillus subtilis encounters phosphate starvation stress, it responds by inducing groups of genes that function to restrict the metabolic consequences of the limited supply of this essential nutrient. These groups of genes are collectively referred to as the phosphate (Pho) stimulon. The phosphate stimulon includes at least two well-described regulons, namely, the sigma B ( B ) general stress regulon and the phosphate starvation-specific PhoP regulon. When B. subtilis encounters phosphate starvation, genes of the SigB regulon are induced by the alternative sigma factor, B , and genes of the PhoP regulon are either induced or repressed by activated PhoP (namely, PhoPϳP). The B general stress regulon contains Ͼ100 genes (58, 64). These genes provide a nonspecific response to stress by encoding proteins that protect the DNA, membranes, and proteins from the damaging effects of stress. Proteins induced by B help the cell to survive potentially harmful environmental conditions, such as heat, osmotic, acid, or alkaline shock (6,21,23,26). This protective function is thought to be particularly important in maintaining the viability of nongrowing cells.The PhoP regulon currently consists of 34 members. Six operons (phoPR [56,60], phoB-ydhF [7,14], pstSAC-pstBApstBB [3,67], phoD-tatAD [7,19], resABCDE [10], and tuaABC DEFGH [40,72]) and five monocistronic genes (glpQ [7], phoA [30,31], tatCD [34], ykoL [60], and yttP [62]) are induced and two operons (tagAB and tagDEF (39) are repressed in response to phosphate starvation. phoA and phoB encode alkaline phosphatases (APases) which facilitate the recovery of inorganic phosphate (P i ) from organic sources (11, 30); phoD encodes a phosphodiesterase/APase, putatively involved in cell wall teichoic acid turnover, and is secreted exclus...

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“…1A). B. subtilis 168 is the best model for the study of teichoic acid biosynthesis as the roles of the enzymes involved in the pathway have been demonstrated biochemically (5,12,27,34) or have been predicted based on strong similarity to archetypal enzymes (21,25,35). This work has given rise to a model for teichoic acid biosynthesis that involves successive elongation of undecaprenyl-linked intermediates, followed by export and attachment to nascent peptidoglycan (Fig.…”

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Use of CDP-Glycerol as an Alternate Acceptor for the Teichoic Acid Polymerase Reveals that Membrane Association Regulates Polymer Length

Schertzer

Brown

2008

J Bacteriol

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The study of bacterial extracellular polysaccharide biosynthesis is hampered by the fact that these molecules are synthesized on membrane-resident carrier lipids. To get around this problem, a practical solution has been to synthesize soluble lipid analogs and study the biosynthetic enzymes using a soluble system. This has been done for the Bacillus subtilis teichoic acid polymerase, TagF, although several aspects of catalysis were inconsistent with the results obtained with reconstituted membrane systems or physiological observations. In this work we explored the acceptor substrate promiscuity and polymer length disregulation that appear to be characteristic of TagF activity away from biological membranes. Using isotope labeling, steady-state kinetics, and chemical lability studies, we demonstrated that the enzyme can synthesize poly(glycerol phosphate) teichoic acid using the elongation substrate CDP-glycerol as an acceptor. This suggests that substrate specificity is relaxed in the region distal to the glycerol phosphate moiety in the acceptor molecule under these conditions. Polymer synthesis proceeded at a rate (27 min ؊1 ) comparable to that in the reconstituted membrane system after a distinct lag period which likely represented slower initiation on the unnatural CDPglycerol acceptor. We confirmed that polymer length became disregulated in the soluble system as the polymers synthesized on CDP-glycerol acceptors were much larger than the polymers synthesized on the membrane or previously found attached to bacterial cell walls. Finally, polymer synthesis on protease-treated membranes suggested that proper length regulation is retained in the absence of accessory proteins and provided evidence that such regulation is conferred through proper association of the polymerase with the membrane.

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Analysis of Bacillus subtilis tag gene expression using transcriptional fusions

Cited by 36 publications

References 45 publications

Teichoic Acid Is an Essential Polymer in Bacillus subtilis That Is Functionally Distinct from Teichuronic Acid

Teichoic Acid Is an Essential Polymer in Bacillus subtilis That Is Functionally Distinct from Teichuronic Acid

Genome-Wide Transcriptional Analysis of the Phosphate Starvation Stimulon ofBacillus subtilis

Use of CDP-Glycerol as an Alternate Acceptor for the Teichoic Acid Polymerase Reveals that Membrane Association Regulates Polymer Length

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