The LacI/GalR family transcriptional regulator UriR negatively controls uridine utilization of Corynebacterium glutamicum by binding to catabolite-responsive element (cre)-like sequences

被引:20
作者
Brinkrolf, Karina [1 ,2 ]
Ploeger, Svenja [1 ]
Solle, Sandra [1 ]
Brune, Iris [1 ]
Nentwich, Svenia S. [1 ,3 ]
Hueser, Andrea T. [1 ]
Kalinowski, Joern [1 ]
Puehler, Alfred [3 ]
Tauch, Andreas [1 ]
机构
[1] Univ Bielefeld, Inst Genomforschung & Systembiol, Ctr Biotechnol, D-33615 Bielefeld, Germany
[2] Univ Bielefeld, Int NRW Grad Sch Bioinformat & Genome Res, Ctr Biotechnol, D-33615 Bielefeld, Germany
[3] Univ Bielefeld, Fak Biol, Lehrstuhl Genet, D-33615 Bielefeld, Germany
来源
MICROBIOLOGY-SGM | 2008年 / 154卷
关键词
D O I
10.1099/mic.0.2007/014001-0
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The Cg1547 protein of Corynebacterium glutamicum ATCC 13032 is a member of the LacI/GalR family of DNA-binding transcriptional regulators. A defined deletion in the cg1547 gene, now designated uriR (uridine utilization regulator), resulted in the mutant strain C. glutamicum KB1547. Comparison of gene expression levels in C. glutamicum KB1547 and the wild-type strain revealed enhanced expression of the uriR operon genes cg1546 (ribokinase), cg1545 (uridine transporter) and cg1543 (uridine-pref erring nucleoside hydrolase). Gene expression of the uriR operon was stimulated by the presence of either uridine or ribose. Growth assays with C. glutamicum mutants showed that functional Cg 1543 and Cg 1545 proteins are essential for the utilization of uridine as the sole carbon source. Transcriptional regulation of the uriR operon is mediated by a 29 bp palindromic sequence composed of two catabolite-responsive element (cre)-like sequences and located in between the mapped -10 promoter region and the start codon of uriR. A similar cre sequence was detected in the upstream region of rbsK2 (cg2554), coding for a second ribokinase in C. glutamicum ATCC 13032. DNA band-shift assays with a streptavidin-tagged UriR protein and labelled oligonucleotides including the cre-like sequences of uriR and rbsK2 demonstrated the specific binding of the purified regulator in vitro. Whole-genome DNA microarray hybridizations comparing the gene expression in C. glutamicum KB1547 with that of the wild-type strain revealed that UriR is a pathway-specific repressor of genes involved in uridine utilization in C. glutamicum.
引用
收藏
页码:1068 / 1081
页数:14
相关论文
共 55 条
[11]   Structural analysis of lac repressor bound to allosteric effectors [J].
Daber, Robert ;
Stayrook, Steven ;
Rosenberg, Allison ;
Lewis, Mitchell .
JOURNAL OF MOLECULAR BIOLOGY, 2007, 370 (04) :609-619
[12]   EMMA:: a platform for consistent storage and efficient analysis of microarray data [J].
Dondrup, M ;
Goesmann, A ;
Bartels, D ;
Kalinowski, J ;
Krause, L ;
Linke, B ;
Rupp, O ;
Sczyrba, A ;
Pühler, A ;
Meyer, F .
JOURNAL OF BIOTECHNOLOGY, 2003, 106 (2-3) :135-146
[13]   Dimerisation mutants of Lac repressor.: I.: A monomeric mutant, L251A, that binds lac operator DNA as a dimer [J].
Dong, FM ;
Spott, S ;
Zimmermann, O ;
Kisters-Woike, B ;
Müller-Hill, B ;
Barker, A .
JOURNAL OF MOLECULAR BIOLOGY, 1999, 290 (03) :653-666
[14]  
Eikmanns B., 2005, HDB CORYNEBACTERIUM, P241
[15]   Parallel evolution of ligand specificity between LacI/GalR family repressors and periplasmic sugar-binding proteins [J].
Fukami-Kobayashi, K ;
Tateno, Y ;
Nishikawa, K .
MOLECULAR BIOLOGY AND EVOLUTION, 2003, 20 (02) :267-277
[16]   DIFFERENTIAL PLASMID RESCUE FROM TRANSGENIC MOUSE DNAS INTO ESCHERICHIA-COLI METHYLATION-RESTRICTION MUTANTS [J].
GRANT, SGN ;
JESSEE, J ;
BLOOM, FR ;
HANAHAN, D .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (12) :4645-4649
[17]   Expression of Corynebacterium glutamicum glycolytic genes varies with carbon source and growth phase [J].
Han, Sung Ok ;
Inui, Masayuki ;
Yukawa, Hideaki .
MICROBIOLOGY-SGM, 2007, 153 :2190-2202
[18]   Reconstruction of microbial transcriptional regulatory networks [J].
Herrgård, MJ ;
Covert, MW ;
Palsson, BO .
CURRENT OPINION IN BIOTECHNOLOGY, 2004, 15 (01) :70-77
[19]   ENGINEERING HYBRID GENES WITHOUT THE USE OF RESTRICTION ENZYMES - GENE-SPLICING BY OVERLAP EXTENSION [J].
HORTON, RM ;
HUNT, HD ;
HO, SN ;
PULLEN, JK ;
PEASE, LR .
GENE, 1989, 77 (01) :61-68
[20]   ANALYSIS OF A CIS-ACTIVE SEQUENCE MEDIATING CATABOLITE REPRESSION IN GRAM-POSITIVE BACTERIA [J].
HUECK, CJ ;
HILLEN, W ;
SAIER, MH .
RESEARCH IN MICROBIOLOGY, 1994, 145 (07) :503-518