Int-B13, an unusual site-specific recombinase of the bacteriophage P4 integrase family, is responsible for chromosomal insertion of the 105-kilobase clc element of Pseudomonas sp. Strain B13.

Détails

ID Serval
serval:BIB_DA6D79C006F5
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Int-B13, an unusual site-specific recombinase of the bacteriophage P4 integrase family, is responsible for chromosomal insertion of the 105-kilobase clc element of Pseudomonas sp. Strain B13.
Périodique
Journal of Bacteriology
Auteur(s)
Ravatn R., Studer S., Zehnder A.J., van der Meer J.R.
ISSN
0021-9193 (Print)
ISSN-L
0021-9193
Statut éditorial
Publié
Date de publication
1998
Peer-reviewed
Oui
Volume
180
Numéro
21
Pages
5505-5514
Langue
anglais
Résumé
Pseudomonas sp. strain B13 carries the clcRABDE genes encoding chlorocatechol-degradative enzymes on the self-transmissible 105-kb clc element. The element integrates site and orientation specifically into the chromosomes of various bacterial recipients, with a glycine tRNA structural gene (glyV) as the integration site. We report here the localization and nucleotide sequence of the integrase gene and the activity of the integrase gene product in mediating site-specific integration. The integrase gene (int-B13) was located near the right end of the clc element. It consisted of an open reading frame (ORF) of maximally 1,971 bp with a coding capacity for 657 amino acids (aa). The full-length protein (74 kDa) was observed upon overexpression and sodium dodecyl sulfate-polyacrylamide gel electrophoresis separation. The N-terminal 430 aa of the predicted Int-B13 protein had substantial similarity to integrases from bacteriophages of the P4 family, but Int-B13 was much larger than P4-type integrases. The C-terminal 220 aa of Int-B13 were homologous to an ORF flanking a gene cluster for naphthalene degradation in Pseudomonas aeruginosa PaK1. Similar to the bacteriophages phiR73 and P4, the clc element integrates into the 3' end of the target tRNA gene. This target site was characterized from four different recipient strains into which the clc element integrated, showing sequence specificity of the integration. In Pseudomonas sp. strain B13, a circular form of the clc element, which carries an 18-bp DNA sequence identical to the 3'-end portion of glyV as part of its attachment site (attP), could be detected. Upon chromosomal integration of the clc element into a bacterial attachment site (attB), a functional glyV was reconstructed at the right end of the element. The integration process could be demonstrated in RecA-deficient Escherichia coli with two recombinant plasmids, one carrying the int-B13 gene and the attP site and the other carrying the attB site of Pseudomonas putida F1.
Mots-clé
Amino Acid Sequence, Base Sequence, Binding Sites, Chromosomes, Bacterial, DNA Nucleotidyltransferases/genetics, DNA Transposable Elements, DNA, Bacterial, Escherichia coli/genetics, Integrases/genetics, Molecular Sequence Data, Pseudomonas Phages/genetics, Pseudomonas putida/genetics, Pseudomonas putida/virology, Recombinases, Sequence Homology, Amino Acid, Virus Integration
Pubmed
Web of science
Création de la notice
21/01/2008 13:36
Dernière modification de la notice
20/08/2019 15:59
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