Identification of a novel steroid inducible gene associated with the ?hsd locus of Comamonas testosteroni

Comamonas testosteroni is a soil bacterium, which can use a variety of steroids as carbon and energy source. Even if it can be estimated that the complete degradation of the steroid nucleus requires more than 20 enzymatic reactions, the complete molecular characterization of the genes encoding these...

Descripción completa

Detalles Bibliográficos
Autores Principales: Pruneda-Paz, José Luis, Linares, Mauricio, Cabrera, Julio E., Genti-Raimondi, Susana
Formato: Artículo (Article)
Lenguaje:Inglés (English)
Publicado: Elsevier 2004
Materias:
Acceso en línea:https://repository.urosario.edu.co/handle/10336/27836
https://doi.org/10.1016/j.jsbmb.2003.10.010
id ir-10336-27836
recordtype dspace
spelling ir-10336-278362021-09-07T21:19:34Z Identification of a novel steroid inducible gene associated with the ?hsd locus of Comamonas testosteroni Identificación de un nuevo gen inducible por esteroides asociado con el locus ?hsd de Comamonas testosteroni Pruneda-Paz, José Luis Linares, Mauricio Cabrera, Julio E. Genti-Raimondi, Susana Steroid Degradation Comamonas testosteroni Comamonas testosteroni is a soil bacterium, which can use a variety of steroids as carbon and energy source. Even if it can be estimated that the complete degradation of the steroid nucleus requires more than 20 enzymatic reactions, the complete molecular characterization of the genes encoding these steroid degradative enzymes as well as the genetic organization of them remain to be elucidated. We have previously reported the cloning and nucleotide sequence of two steroid-inducible genes, ?hsd and stdC encoding 3?-17?-hydroxysteroid dehydrogenase and a hypothetical protein respectively, located in both ends of a 3.2 kb HindIII fragment. Herein, we report the cloning and characterization of another steroid-inducible gene, called sip48 (steroid inducible protein), located between these two genes. The analysis of Sip48 amino acid sequence predicts a protein of 438 amino acids with a molecular mass of 48.5 kDa. This protein bears high homology with conserved hypothetical proteins of unknown function described in Pseudomonas aeruginosa, Pseudomonas syringae, Pseudomonas putida, Burkholderia fungorum, Shewanella oneidensis, Pseudomonas fluorescens and Thauera aromatica. The predicted protein shows a typical structure of a leader peptide at its N-terminus. A 48.5 kDa protein encoded by the recombinant plasmid was detected by SDS–PAGE analysis of in vitro []-methionine labeled polypeptides. Analysis of gene expression indicates that Sip48 is tightly controlled at the transcriptional level by several steroid compounds. In addition, transcriptional analysis of sip48 and ?hsd in a sip48 mutant strain, indicates that both genes are transcribed as a polycistronic mRNA. lacZ transcriptional fusions integrated into the chromosome of C. testosteroni demonstrate that a steroid-inducible promoter located upstream of sip48 regulates the expression of both genes. 2004-01-01 2020-08-19T14:44:09Z info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion ISSN: 0960-0760 EISSN: 1879-1220 https://repository.urosario.edu.co/handle/10336/27836 https://doi.org/10.1016/j.jsbmb.2003.10.010 eng info:eu-repo/semantics/restrictedAccess application/pdf Elsevier Journal of Steroid Biochemistry and Molecular Biology
institution EdocUR - Universidad del Rosario
collection DSpace
language Inglés (English)
topic Steroid
Degradation
Comamonas testosteroni
spellingShingle Steroid
Degradation
Comamonas testosteroni
Pruneda-Paz, José Luis
Linares, Mauricio
Cabrera, Julio E.
Genti-Raimondi, Susana
Identification of a novel steroid inducible gene associated with the ?hsd locus of Comamonas testosteroni
description Comamonas testosteroni is a soil bacterium, which can use a variety of steroids as carbon and energy source. Even if it can be estimated that the complete degradation of the steroid nucleus requires more than 20 enzymatic reactions, the complete molecular characterization of the genes encoding these steroid degradative enzymes as well as the genetic organization of them remain to be elucidated. We have previously reported the cloning and nucleotide sequence of two steroid-inducible genes, ?hsd and stdC encoding 3?-17?-hydroxysteroid dehydrogenase and a hypothetical protein respectively, located in both ends of a 3.2 kb HindIII fragment. Herein, we report the cloning and characterization of another steroid-inducible gene, called sip48 (steroid inducible protein), located between these two genes. The analysis of Sip48 amino acid sequence predicts a protein of 438 amino acids with a molecular mass of 48.5 kDa. This protein bears high homology with conserved hypothetical proteins of unknown function described in Pseudomonas aeruginosa, Pseudomonas syringae, Pseudomonas putida, Burkholderia fungorum, Shewanella oneidensis, Pseudomonas fluorescens and Thauera aromatica. The predicted protein shows a typical structure of a leader peptide at its N-terminus. A 48.5 kDa protein encoded by the recombinant plasmid was detected by SDS–PAGE analysis of in vitro []-methionine labeled polypeptides. Analysis of gene expression indicates that Sip48 is tightly controlled at the transcriptional level by several steroid compounds. In addition, transcriptional analysis of sip48 and ?hsd in a sip48 mutant strain, indicates that both genes are transcribed as a polycistronic mRNA. lacZ transcriptional fusions integrated into the chromosome of C. testosteroni demonstrate that a steroid-inducible promoter located upstream of sip48 regulates the expression of both genes.
format Artículo (Article)
author Pruneda-Paz, José Luis
Linares, Mauricio
Cabrera, Julio E.
Genti-Raimondi, Susana
author_facet Pruneda-Paz, José Luis
Linares, Mauricio
Cabrera, Julio E.
Genti-Raimondi, Susana
author_sort Pruneda-Paz, José Luis
title Identification of a novel steroid inducible gene associated with the ?hsd locus of Comamonas testosteroni
title_short Identification of a novel steroid inducible gene associated with the ?hsd locus of Comamonas testosteroni
title_full Identification of a novel steroid inducible gene associated with the ?hsd locus of Comamonas testosteroni
title_fullStr Identification of a novel steroid inducible gene associated with the ?hsd locus of Comamonas testosteroni
title_full_unstemmed Identification of a novel steroid inducible gene associated with the ?hsd locus of Comamonas testosteroni
title_sort identification of a novel steroid inducible gene associated with the ?hsd locus of comamonas testosteroni
publisher Elsevier
publishDate 2004
url https://repository.urosario.edu.co/handle/10336/27836
https://doi.org/10.1016/j.jsbmb.2003.10.010
_version_ 1712098631623901184
score 12,131701