HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Manzanera, M. Articles by Marqués, S. Search for Related Content PubMed PubMed Citation Articles by Manzanera, M. Articles by Marqués, S. Agricola Articles by Manzanera, M. Articles by Marqués, S.

Molecular characterization of Pseudomonas putida KT2440 rpoH gene regulation

Estación Experimental del Zaidín, CSIC, Departamento de Bioquímica y Biología Molecular y Celular de Plantas, Apartado 419, E-18080 Granada, Spain¹

Author for correspondence: Silvia Marqués. Tel: +34 958 121011. Fax: +34 958 129600. e-mail: silvia{at}eez.csic.es

The rpoH gene of Pseudomonas putida KT2440 encoding the heat-shock sigma factor ³² was cloned and sequenced, and the translated gene product was predicted to be a protein of 32·5 kDa. The unambiguous role of the gene as a sigma factor was confirmed because the cloned P. putida gene complemented the growth defect, at 37 and 42 °C, of an Escherichia coli rpoH mutant strain. Primer extension analysis showed that in P. putida the rpoH gene is expressed from three promoters in cells growing at 30 °C. Two of them, P1 and P3, share homology with the ⁷⁰-dependent promoters, while the third one, P2, shows a typical ²⁴-consensus sequence. The pattern of transcription initiation of the rpoH gene did not change in response to different stresses, i.e. a sudden heat shock or the addition of aromatic compounds. However, the predicted secondary structure of the 5' region of the mRNA derived from the three different promoters suggests regulation at the level of translation efficiency and/or mRNA half-life. An inverted repeat sequence located 20 bp downstream of the rpoH stop codon was shown to function as a terminator in vivo in P. putida growing at temperatures from 18 to 42 °C.

Keywords: ³², stress response, aromatic hydrocarbons, heat-shock response, downstream box

Abbreviations: DEPC, diethyl pyrocarbonate; DS box, downstream box; HS response, heat-shock response

The GenBank accession number for the sequence corresponding to the P. putida KT2440 chromosomal DNA fragment (2891 bp) cloned in pSH27, including the rpoH gene, is AF157048.

This article has been cited by other articles:

				M. Miyakoshi, M. Shintani, T. Terabayashi, S. Kai, H. Yamane, and H. Nojiri Transcriptome Analysis of Pseudomonas putida KT2440 Harboring the Completely Sequenced IncP-7 Plasmid pCAR1 J. Bacteriol., October 1, 2007; 189(19): 6849 - 6860. [Abstract] [Full Text] [PDF]

				P. Dominguez-Cuevas, P. Marin, J. L. Ramos, and S. Marques RNA Polymerase Holoenzymes Can Share a Single Transcription Start Site for the Pm Promoter: CRITICAL NUCLEOTIDES IN THE -7 TO-18 REGION ARE NEEDED TO SELECT BETWEEN RNA POLYMERASE WITH {sigma}38 OR {sigma}32 J. Biol. Chem., December 16, 2005; 280(50): 41315 - 41323. [Abstract] [Full Text] [PDF]

				M.-M. Gonzalez-Perez, J. L. Ramos, and S. Marques Cellular XylS Levels Are a Function of Transcription of xylS from Two Independent Promoters and the Differential Efficiency of Translation of the Two mRNAs J. Bacteriol., March 15, 2004; 186(6): 1898 - 1901. [Abstract] [Full Text] [PDF]

				I. Aranda-Olmedo, R. Tobes, M. Manzanera, J. L. Ramos, and S. Marques Species-specific repetitive extragenic palindromic (REP) sequences in Pseudomonas putida Nucleic Acids Res., April 15, 2002; 30(8): 1826 - 1833. [Abstract] [Full Text] [PDF]