Novel physiological modulation of the Pu promoter of TOL plasmid -: Negative regulatory role of the TurA protein of Pseudomonas putida in the response to suboptimal growth temperatures

被引:37
作者
Rescalli, E
Saini, S
Bartocci, C
Rychlewski, L
de Lorenzo, V
Bertoni, G
机构
[1] Univ Milan, Dipartimento Sci Biomol & Biotecnol, I-20133 Milan, Italy
[2] BioInfoBank Inst, PL-60744 Poznan, Poland
[3] CSIC, Ctr Nacl Biotecnol, Dept Microbial Biotechnol, E-28049 Madrid, Spain
关键词
D O I
10.1074/jbc.M310580200
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
From crude protein extracts of Pseudomonas putida KT2440, we identified a small protein, TurA, able to bind to DNA fragments bearing the entire Pu promoter sequence of the TOL plasmid. The knock-out inactivation of the turA gene resulted in enhanced transcription initiation from the Pu promoter, initially suggesting a negative regulatory role of TurA on Pu expression. Ectopic expression of TurA both in P. putida and in Escherichia coli reporter strains and transcription in vitro of the Pu promoter in the presence of purified TurA confirmed the TurA repressor role on Pu activity. turA gene inactivation did not significantly alter two well characterized physiological regulations of the Pu expression in routine conditions of cultivation, exponential silencing, and carbon-mediated repression, respectively. However, the growth at suboptimal temperatures resulted in a TurA-dependent increase of Pu repression. These results strongly suggest that a physiological significance of the negative role of TurA on Pu activity could be limitation of the expression of the toluene-degrading enzymes at suboptimal growth temperatures. Therefore, the identification of TurA as Pu-binding protein revealed a novel physiological modulation of Pu promoter that is different from those strictly nutritional described previously.
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收藏
页码:7777 / 7784
页数:8
相关论文
共 57 条
[1]   Gapped BLAST and PSI-BLAST: a new generation of protein database search programs [J].
Altschul, SF ;
Madden, TL ;
Schaffer, AA ;
Zhang, JH ;
Zhang, Z ;
Miller, W ;
Lipman, DJ .
NUCLEIC ACIDS RESEARCH, 1997, 25 (17) :3389-3402
[2]   BASIC LOCAL ALIGNMENT SEARCH TOOL [J].
ALTSCHUL, SF ;
GISH, W ;
MILLER, W ;
MYERS, EW ;
LIPMAN, DJ .
JOURNAL OF MOLECULAR BIOLOGY, 1990, 215 (03) :403-410
[3]   H-NS: A modulator of environmentally regulated gene expression [J].
Atlung, T ;
Ingmer, H .
MOLECULAR MICROBIOLOGY, 1997, 24 (01) :7-17
[4]  
Bateman A, 2004, NUCLEIC ACIDS RES, V32, pD138, DOI [10.1093/nar/gkp985, 10.1093/nar/gkr1065, 10.1093/nar/gkh121]
[5]   H-NS and H-NS-like proteins in Gram-negative bacteria and their multiple role in the regulation of bacterial metabolism [J].
Bertin, P ;
Hommais, F ;
Krin, E ;
Soutourina, O ;
Tendeng, C ;
Derzelle, S ;
Danchin, A .
BIOCHIMIE, 2001, 83 (02) :235-241
[6]   Active recruitment of σ54 RNA polymerase to the Pu promoter of Pseudomonas putida:: role of IHF and αCTD [J].
Bertoni, G ;
Fujita, N ;
Ishihama, A ;
de Lorenzo, V .
EMBO JOURNAL, 1998, 17 (17) :5120-5128
[7]   The bacterial enhancer-dependent σ54 (σN) transcription factor [J].
Buck, M ;
Gallegos, MT ;
Studholme, DJ ;
Guo, YL ;
Gralla, JD .
JOURNAL OF BACTERIOLOGY, 2000, 182 (15) :4129-4136
[8]   Recruitment of RNA polymerase is a rate-limiting step for the activation of the σ54 promoter Pu of Pseudomonas putida [J].
Carmona, M ;
de Lorenzo, V ;
Bertoni, G .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (47) :33790-33794
[9]   Expression systems and physiological control of promoter activity in bacteria [J].
Cases, I ;
de Lorenzo, V .
CURRENT OPINION IN MICROBIOLOGY, 1998, 1 (03) :303-310
[10]   The IIANtr (PtsN) protein of Pseudomonas putida mediates the C source inhibition of the σ54-dependent Pu promoter of the TOL plasmid [J].
Cases, I ;
Pérez-Martín, J ;
de Lorenzo, V .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (22) :15562-15568