Regulation of the central glycolytic genes in Bacillus subtilis:: binding of the repressor CggR to its single DNA target sequence is modulated by fructose-1,6-bisphosphate

被引:79
作者
Doan, T
Aymerich, S [1 ]
机构
[1] INRA, UMR216, CNRS, URA1925, F-78850 Thiverval Grignon, France
[2] INAPG, F-78850 Thiverval Grignon, France
关键词
D O I
10.1046/j.1365-2958.2003.03404.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Glycolysis is one of the best and widely conserved general metabolic pathways. Bacillus subtilis enzymes catalysing the central part of glycolysis, gathering the steps of interconversion of the triose phosphates from dihydroxyacetone-phosphate to phosphoenolpyruvate, are encoded by five genes, gapA, pgk, tpi, pgm and eno. They are transcribed in a hexacistronic operon together with cggR , the first cistron, encoding the repressor of this gapA operon. Using deletion analysis, we have localized the CggR operator between the promoter and the first gene of the operon. CggR was purified and used in gel mobility shift assays and DNase I footprinting experiments to delimit its target sequence. Site-directed mutagenesis and in vivo tests demonstrated that it consists of two direct-repeats (CGGGACN(6)TGTCN(4)CGGGACN(6)TGTC). Sequence analysis and transcriptome comparison of a wild-type and a cggR mutant strain strongly suggested that CggR regulates only the gapA operon. The presence of glycolytic carbon sources induces expression of the gapA operon. Genetic experiments allowed us to identify the metabolic steps required for the formation of the CggR effector. In vitro experiments with the suggested candidates allowed us to demonstrate that fructose-1,6-biphosphate (FBP) acts as an inhibitor of CggR DNA-binding activity (10 mM for full inhibition). FBP is thus the major signal for both CcpA-dependent catabolite repression (or activation) and activation of the central glycolytic genes. Genomic sequence comparisons suggest that these results can apply to numerous low-G+C, Gram-positive bacterial species.
引用
收藏
页码:1709 / 1721
页数:13
相关论文
共 51 条
[21]   Crystal structure of the transcription activator BmrR bound to DNA and a drug [J].
Heldwein, EEZ ;
Brennan, RG .
NATURE, 2001, 409 (6818) :378-382
[22]  
Henkin TM, 1996, FEMS MICROBIOL LETT, V135, P9
[23]   Genes for D-arabinitol and ribitol catabolism from Klebsiella pneumoniae [J].
Heuel, H ;
Shakeri-Garakani, A ;
Turgut, S ;
Lengeler, JW .
MICROBIOLOGY-SGM, 1998, 144 :1631-1639
[24]   CATABOLITE REPRESSION IN BACILLUS-SUBTILIS - A GLOBAL REGULATORY MECHANISM FOR THE GRAM-POSITIVE BACTERIA [J].
HUECK, CJ ;
HILLEN, W .
MOLECULAR MICROBIOLOGY, 1995, 15 (03) :395-401
[25]   Prokaryotic transcription regulators: more than just the helix-turn-helix motif [J].
Huffman, JL ;
Brennan, RG .
CURRENT OPINION IN STRUCTURAL BIOLOGY, 2002, 12 (01) :98-106
[26]   The complete genome sequence of the Gram-positive bacterium Bacillus subtilis [J].
Kunst, F ;
Ogasawara, N ;
Moszer, I ;
Albertini, AM ;
Alloni, G ;
Azevedo, V ;
Bertero, MG ;
Bessieres, P ;
Bolotin, A ;
Borchert, S ;
Borriss, R ;
Boursier, L ;
Brans, A ;
Braun, M ;
Brignell, SC ;
Bron, S ;
Brouillet, S ;
Bruschi, CV ;
Caldwell, B ;
Capuano, V ;
Carter, NM ;
Choi, SK ;
Codani, JJ ;
Connerton, IF ;
Cummings, NJ ;
Daniel, RA ;
Denizot, F ;
Devine, KM ;
Dusterhoft, A ;
Ehrlich, SD ;
Emmerson, PT ;
Entian, KD ;
Errington, J ;
Fabret, C ;
Ferrari, E ;
Foulger, D ;
Fritz, C ;
Fujita, M ;
Fujita, Y ;
Fuma, S ;
Galizzi, A ;
Galleron, N ;
Ghim, SY ;
Glaser, P ;
Goffeau, A ;
Golightly, EJ ;
Grandi, G ;
Guiseppi, G ;
Guy, BJ ;
Haga, K .
NATURE, 1997, 390 (6657) :249-256
[27]   CLONING AND NUCLEOTIDE-SEQUENCES OF THE GENES ENCODING TRIOSE PHOSPHATE ISOMERASE, PHOSPHOGLYCERATE MUTASE, AND ENOLASE FROM BACILLUS-SUBTILIS [J].
LEYVAVAZQUEZ, MA ;
SETLOW, P .
JOURNAL OF BACTERIOLOGY, 1994, 176 (13) :3903-3910
[28]   Control of the glycolytic gapA operon by the catabolite control protein A in Bacillus subtilis:: a novel mechanism of CcpA-mediated regulation [J].
Ludwig, H ;
Rebhan, N ;
Blencke, HM ;
Merzbacher, M ;
Stülke, J .
MOLECULAR MICROBIOLOGY, 2002, 45 (02) :543-553
[29]   Transcription of glycolytic genes and operons in Bacillus subtilis:: evidence for the presence of multiple levels of control of the gapA operon [J].
Ludwig, H ;
Homuth, G ;
Schmalisch, M ;
Dyka, FM ;
Hecker, M ;
Stülke, J .
MOLECULAR MICROBIOLOGY, 2001, 41 (02) :409-422
[30]   Pyrophosphate-producing protein dephosphorylation by HPr kinase/phosphorylase: A relic of early life? [J].
Mijakovic, I ;
Poncet, S ;
Galinier, A ;
Monedero, V ;
Fieulaine, S ;
Janin, J ;
Nessler, S ;
Marquez, JA ;
Scheffzek, K ;
Hasenbein, S ;
Hengstenberg, W ;
Deutscher, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (21) :13442-13447