The switch from inorganic to organic sulphur assimilation in Escherichia coli:: adenosine 5′-phosphosulphate (APS) as a signalling molecule for sulphate excess

被引:48
作者
Bykowski, T
van der Ploeg, JR
Iwanicka-Nowicka, R
Hryniewicz, MM
机构
[1] Polish Acad Sci, Inst Biochem & Biophys, PL-02106 Warsaw, Poland
[2] ETH Zentrum, Swiss Fed Inst Technol, Inst Microbiol, CH-8092 Zurich, Switzerland
关键词
D O I
10.1046/j.1365-2958.2002.02846.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
The utilization of organosulphur compounds as sources of sulphur by Escherichia coli is strongly repressed by sulphate. To search for the signal enabling E. coli to alternate gene expression according to the sulphur source, we investigated the transcriptional control of the ssuEADCB operon, required for the transport and desulphonation of aliphatic sulphonates. We demonstrate that, of the two LysR-type regulators involved in expression from the ssu promoter, CbI acts as a direct and sufficient activator of transcription in vivo and in vitro, whereas CysB downregulates the promoter efficiency. Most importantly, the Cbl-medlated transcription initiation at the ssu promoter in vitro is abolished in the presence of an early metabolite of the sulphate assimilatory pathway, adenosine 5'-phosphosulphate (APS). This role for APS was confirmed In vivo by measuring the expression of beta-galactosidase from a transcriptional ssu-lacZ fusion in strains containing different mutations blocking the synthesis and consumption of APS. Our data comprise the first evidence that APS may act as the negative cofactor of the transcriptional regulator Cbl, and that APS, and not sulphate itself, serves as the signalling molecule for sulphate excess.
引用
收藏
页码:1347 / 1358
页数:12
相关论文
共 37 条
[1]  
Ausubel FM., 1994, Curr. Protoc. Mol. Biol
[2]   Transcription activation at the Escherichia coli melAB promoter:: the role of MelR and the cyclic AMP receptor protein [J].
Belyaeva, TA ;
Wade, JT ;
Webster, CL ;
Howard, VJ ;
Thomas, MS ;
Hyde, EI ;
Busby, SJW .
MOLECULAR MICROBIOLOGY, 2000, 36 (01) :211-222
[3]   RESIDUE THREONINE-149 OF THE SALMONELLA-TYPHIMURIUM CYSB TRANSCRIPTION ACTIVATOR - MUTATIONS CAUSING CONSTITUTIVE EXPRESSION OF POSITIVELY REGULATED GENES OF THE CYSTEINE REGULON [J].
COLYER, TE ;
KREDICH, NM .
MOLECULAR MICROBIOLOGY, 1994, 13 (05) :797-805
[4]   In vitro characterization of constitutive CysB proteins from Salmonella typhimurium [J].
Colyer, TE ;
Kredich, NM .
MOLECULAR MICROBIOLOGY, 1996, 21 (02) :247-256
[5]   Microbial desulfonation [J].
Cook, AM ;
Laue, H ;
Junker, F .
FEMS MICROBIOLOGY REVIEWS, 1998, 22 (05) :399-419
[6]   One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products [J].
Datsenko, KA ;
Wanner, BL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (12) :6640-6645
[7]   Characterization of alpha-ketoglutarate-dependent taurine dioxygenase from Escherichia coli [J].
Eichhorn, E ;
vanderPloeg, JR ;
Kertesz, MA ;
Leisinger, T .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (37) :23031-23036
[8]   Characterization of a two-component alkanesulfonate monooxygenase from Escherichia coli [J].
Eichhorn, E ;
van der Ploeg, JR ;
Leisinger, T .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (38) :26639-26646
[9]   CONSERVED MOTIFS IN A DIVERGENT NOD BOX OF AZORHIZOBIUM-CAULINODANS ORS571 REVEAL A COMMON STRUCTURE IN PROMOTERS REGULATED BY LYSR-TYPE PROTEINS [J].
GOETHALS, K ;
VANMONTAGU, M ;
HOLSTERS, M .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (05) :1646-1650
[10]   THE CYSP PROMOTER OF SALMONELLA-TYPHIMURIUM - CHARACTERIZATION OF 2 BINDING-SITES FOR CYSB PROTEIN, STUDIES OF INVIVO TRANSCRIPTION INITIATION, AND DEMONSTRATION OF THE ANTI-INDUCER EFFECTS OF THIOSULFATE [J].
HRYNIEWICZ, MM ;
KREDICH, NM .
JOURNAL OF BACTERIOLOGY, 1991, 173 (18) :5876-5886