Induction of Levansucrase in Bacillus Subtilis: an Antitermination Mechanism Negatively Controlled by the Phosphotransferase System

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1990 Feb 1

PMID 2105292

Citations 51

Authors

A M Crutz

M Steinmetz

S Aymerich

R Richter

D Le Coq

Affiliations

Soon will be listed here.

Abstract

The target of the induction by sucrose of the levansucrase gene is a transcription terminator (sacRt) located upstream from the coding sequence, sacB. The two-gene locus sacX-sacY (formerly sacS) and the ptsI gene were previously shown to be involved in this induction. ptsI encodes enzyme I of the phosphoenolpyruvate-dependent phosphotransferase system. SacX is strongly homologous to sucrose-specific phosphotransferase system-dependent permeases. SacY is a positive regulator of sacB. Here we show that SacY is probably an antiterminator interacting directly with sacRt, since in Escherichia coli the presence of the sacY gene stimulates the expression of a reporter gene fused downstream from sacRt. Missense mutations affecting sacY were sequenced, and the sacB regulation was studied in isogenic strains carrying these mutations or in vitro-generated mutations affecting sacX, sacY, or ptsI. The phenotype of double mutants suggests a model in which SacX might be a sucrose sensor that would be phosphorylated by the phosphotransferase system and, in this state, could inhibit the SacY antiterminator. Exogenous sucrose, or a mutation inactivating the phosphotransferase system, would dephosphorylate SacX and allow antitermination at sacRt.

Citing Articles

Combinatorial metabolic engineering enables high yield production of α-arbutin from sucrose by biocatalysis.

Zhou Q, Wu Y, Deng J, Liu Y, Li J, Du G Appl Microbiol Biotechnol. 2023; 107(9):2897-2910.

PMID: 37000229 DOI: 10.1007/s00253-023-12496-2.

Influence of the phosphoenolpyruvate:carbohydrate phosphotransferase system on toxin gene expression and virulence in Bacillus anthracis.

Bier N, Hammerstrom T, Koehler T Mol Microbiol. 2019; 113(1):237-252.

PMID: 31667937 PMC: 8462991. DOI: 10.1111/mmi.14413.

Cross Talk among Transporters of the Phosphoenolpyruvate-Dependent Phosphotransferase System in Bacillus subtilis.

Heravi K, Altenbuchner J J Bacteriol. 2018; 200(19).

PMID: 30038046 PMC: 6148471. DOI: 10.1128/JB.00213-18.

Genome engineering using a synthetic gene circuit in Bacillus subtilis.

Jeong D, Park S, Pan J, Kim E, Choi S Nucleic Acids Res. 2015; 43(6):e42.

PMID: 25552415 PMC: 4381049. DOI: 10.1093/nar/gku1380.

Translation efficiency of antiterminator proteins is a determinant for the difference in glucose repression of two β-glucoside phosphotransferase system gene clusters in Corynebacterium glutamicum R.

Tanaka Y, Teramoto H, Inui M, Yukawa H J Bacteriol. 2010; 193(2):349-57.

PMID: 21075922 PMC: 3019825. DOI: 10.1128/JB.01123-10.

References

Lepesant J, Kunst F, DEDONDER R . Chromosomal location of mutations affecting sucrose metabolism in Bacillus subtilis Marburg. Mol Gen Genet. 1972; 118(2):135-60. DOI: 10.1007/BF00267084. View

Postma P, Lengeler J . Phosphoenolpyruvate:carbohydrate phosphotransferase system of bacteria. Microbiol Rev. 1985; 49(3):232-69. PMC: 373035. DOI: 10.1128/mr.49.3.232-269.1985. View

SANGER F, Nicklen S, Coulson A . DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977; 74(12):5463-7. PMC: 431765. DOI: 10.1073/pnas.74.12.5463. View

Chang A, Cohen S . Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978; 134(3):1141-56. PMC: 222365. DOI: 10.1128/jb.134.3.1141-1156.1978. View

Kadner R . Exogenous induction of the Escherichia coli hexose phosphate transport system defined by uhp-lac operon fusions. J Bacteriol. 1981; 148(1):203-9. PMC: 216182. DOI: 10.1128/jb.148.1.203-209.1981. View

Horinouchi S, Weisblum B . Nucleotide sequence and functional map of pE194, a plasmid that specifies inducible resistance to macrolide, lincosamide, and streptogramin type B antibodies. J Bacteriol. 1982; 150(2):804-14. PMC: 216433. DOI: 10.1128/jb.150.2.804-814.1982. View

FERRARI F, Ferrari E, Hoch J . Chromosomal location of a Bacillus subtilis DNA fragment uniquely transcribed by sigma-28-containing RNA polymerase. J Bacteriol. 1982; 152(2):780-5. PMC: 221529. DOI: 10.1128/jb.152.2.780-785.1982. View

Weston L, Kadner R . Role of uhp genes in expression of the Escherichia coli sugar-phosphate transport system. J Bacteriol. 1988; 170(8):3375-83. PMC: 211304. DOI: 10.1128/jb.170.8.3375-3383.1988. View

de Reuse H, Danchin A . The ptsH, ptsI, and crr genes of the Escherichia coli phosphoenolpyruvate-dependent phosphotransferase system: a complex operon with several modes of transcription. J Bacteriol. 1988; 170(9):3827-37. PMC: 211377. DOI: 10.1128/jb.170.9.3827-3837.1988. View

10.

Schnetz K, Rak B . Regulation of the bgl operon of Escherichia coli by transcriptional antitermination. EMBO J. 1988; 7(10):3271-7. PMC: 454751. DOI: 10.1002/j.1460-2075.1988.tb03194.x. View

11.

Eisermann R, Deutscher J, Hengstenberg W . Site-directed mutagenesis with the ptsH gene of Bacillus subtilis. Isolation and characterization of heat-stable proteins altered at the ATP-dependent regulatory phosphorylation site. J Biol Chem. 1988; 263(32):17050-4. View

12.

Sloma A, Ally A, Ally D, Pero J . Gene encoding a minor extracellular protease in Bacillus subtilis. J Bacteriol. 1988; 170(12):5557-63. PMC: 211651. DOI: 10.1128/jb.170.12.5557-5563.1988. View

13.

Kawamura F, Wang L, Doi R . Catabolite-resistant sporulation (crsA) mutations in the Bacillus subtilis RNA polymerase sigma 43 gene (rpoD) can suppress and be suppressed by mutations in spo0 genes. Proc Natl Acad Sci U S A. 1985; 82(23):8124-8. PMC: 391455. DOI: 10.1073/pnas.82.23.8124. View

14.

Aymerich S, Steinmetz M . 5'-noncoding region sacR is the target of all identified regulation affecting the levansucrase gene in Bacillus subtilis. J Bacteriol. 1986; 166(3):993-8. PMC: 215223. DOI: 10.1128/jb.166.3.993-998.1986. View

15.

Shimotsu H, Henner D . Construction of a single-copy integration vector and its use in analysis of regulation of the trp operon of Bacillus subtilis. Gene. 1986; 43(1-2):85-94. DOI: 10.1016/0378-1119(86)90011-9. View

16.

Shimotsu H, Henner D . Modulation of Bacillus subtilis levansucrase gene expression by sucrose and regulation of the steady-state mRNA level by sacU and sacQ genes. J Bacteriol. 1986; 168(1):380-8. PMC: 213462. DOI: 10.1128/jb.168.1.380-388.1986. View

17.

Steinmetz M . Phosphoenolpyruvate:sugar phosphotransferase system of Bacillus subtilis: cloning of the region containing the ptsH and ptsI genes and evidence for a crr-like gene. J Bacteriol. 1987; 169(5):2287-90. PMC: 212157. DOI: 10.1128/jb.169.5.2287-2290.1987. View

18.

Ronson C, Nixon B, Ausubel F . Conserved domains in bacterial regulatory proteins that respond to environmental stimuli. Cell. 1987; 49(5):579-81. DOI: 10.1016/0092-8674(87)90530-7. View

19.

Steinmetz M, Le Coq D, Djemia H, Gay P . [Genetic analysis of sacB, the structural gene of a secreted enzyme, levansucrase of Bacillus subtilis Marburg]. Mol Gen Genet. 1983; 191(1):138-44. DOI: 10.1007/BF00330901. View

20.

Mahadevan S, Reynolds A, Wright A . Positive and negative regulation of the bgl operon in Escherichia coli. J Bacteriol. 1987; 169(6):2570-8. PMC: 212126. DOI: 10.1128/jb.169.6.2570-2578.1987. View