Abstract
Plasmid pRJ9 is a non-self-mobilizable bacteriocinogenic plasmid from Staphylococcus aureus. Despite this feature, DNA sequencing and RT-PCR experiments showed that it presents a Mob region with three genes (mobCAB), transcribed as an operon. In silico analysis of the Mob proteins encoded by pRJ9 showed that they present all the conserved functional features reported until present as being essential for plasmid mobilization. Moreover, they showed a high identity to Mob proteins encoded by mobilizable plasmids from Staphylococcus spp., especially to those encoded by plasmid pRJ6, which presents four mob genes (mobCDAB). A putative oriT region was also found upstream of the pRJ9 mob operon. pRJ9 could only be successfully mobilized by pGO1 when pRJ6 was present in the same strain. Further experiments showed that the pRJ9 oriT can be recognized by the pRJ6 Mob proteins, confirming its functionality. As pRJ9 does not possess a mobD gene while pRJ6 does, the absence of this gene was believed to be responsible for its lack of mobilization. However, conjugation experiments with a donor strain carrying also mobD cloned into an S. aureus vector showed that pRJ9 does not become mobilized even in the presence of the protein MobD encoded by pRJ6. Therefore, the reasons for pRJ9 failure to be mobilized are presently unknown.
Journal Section:
Research Article
References
1.
Abajy MY, Kopeć J, Schiwon K, Burzynski M, Döring M, Bohn C, Grohmann E: A type IV-secretion-like system is required for conjugative DNA transport of broad-host-range plasmid pIP501 in Gram-positive bacteria. J Bacteriol 2007;189:2487–2496.
[PubMed]
2.
Apisiridej S, Leelaporn A, Scaramuzzi CD, Skurray RA, Firth N: Molecular analysis of a mobilizable theta-mode trimethoprim resistance plasmid from coagulase-negative staphylococci. Plasmid 1997;38:13–24.
[PubMed]
3.
Augustin J, Rosenstein R, Wieland B, Schneider U, Schnell N, Engelke G, Entian KD, Götz F: Genetic analysis of epidermin biosynthetic genes and epidermin-negative mutants of Staphylococcus epidermidis. Eur J Biochem 1992;204:1149–1154.
[PubMed]
4.
Backert S, Meyer TF: Type IV secretion systems and their effectors in bacterial pathogenesis. Curr Opin Microbiol 2006;9:207–217.
[PubMed]
5.
Bastos MCF, Bonaldo MC, Penido EG: Constitutive erythromycin resistance plasmid in Staphylococcus aureus. J Gen Microbiol 1980;121:513–516.
[PubMed]
6.
Caryl JA, Smith MCA, Thomas CD: Reconstitution of a staphylococcal plasmid-protein complex in vitro. J Bacteriol 2004;186:3374–3383.
[PubMed]
7.
Caryl JA, Thomas CD: Investigating the basis of substrate recognition in the pC221 relaxosome. Mol Microbiol 2006;60:1302–1318.
[PubMed]
8.
Charpentier E, Anton AI, Barry P, Alfonso B, Fang Y, Novick RP: Novel cassette-based shuttle vector system for Gram-positive bacteria. Appl Environ Microbiol 2004;70:6076–6085.
[PubMed]
9.
Coelho MLV, Ceotto H, Madureira DJ, Nes IF, Bastos MCF: Mobilization functions of the bacteriocinogenic plasmid pRJ6 of Staphylococcus aureus. J Microbiol 2009;47:327–336.
[PubMed]
10.
Francia MV, Varsaki A, Garcillán-Barcia MP, LaTorre A, Drainas C, de la Cruz F: A classification scheme for mobilization regions of bacterial plasmids. FEMS Microbiol Rev 2004;28:79–100.
[PubMed]
11.
Gamon MR, Moreira EC, Oliveira SS, Teixeira LM, Bastos MCF: Characterization of a novel bacteriocin-encoding plasmid found in clinical isolates of Staphylococcus aureus. Antonie van Leeuwenhoek 1999;75:233–243.
12.
Giambiagi-deMarval M, Mafra MA, Penido EGC, Bastos MCF: Distinct groups of plasmids correlated with bacteriocin production in Staphylococcus aureus. J Gen Microbiol 1990;136:1591–1599.
[PubMed]
13.
Grohmann E, Muth G, Espinosa M: Conjugative plasmid transfer in Gram-positive bacteria. Microbiol. Mol Biol Rev 2003;67:277–301.
[PubMed]
14.
Netz DJA, Pohl R, Beck-Sickinger AG, Selmer T, Pierik AJ, Bastos MCF, Sahl H-G: Biochemical characterisation and genetic analysis of aureocin A53, a new, atypical bacteriocin from Staphylococcus aureus. J Mol Biol 2002;319:745–756.
[PubMed]
15.
Novick RP: Properties of a cryptic high-frequency transducing phage in Staphylococcus aureus. Virology 1967;33:155–166.
[PubMed]
16.
Oliveira SS, Nascimento JS, Póvoa DC, Araújo SA, Gamon MR, Bastos MCF: Genetic analysis of the bacteriocin-encoding plasmids pRJ6 and pRJ9 of Staphylococcus aureus by transposon mutagenesis and cloning of genes involved in bacteriocin production. J Appl Microbiol 1998;85:972–984.
[PubMed]
17.
Projan SJ, Archer GL: Mobilization of the relaxable Staphylococcus aureus plasmid pC221 by the conjugative plasmid pGOl involves three pC221 loci. J Bacteriol 1989;171:1841–1845.
[PubMed]
18.
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, ed 2. New York, Cold Spring Harbor Laboratory Press, 1989.
19.
Schenk S, Laddaga RA: Improved method for electroporation of Staphylococcus aureus. FEMS Microbiol Lett 1992;73:133–138.
[PubMed]
20.
Smillie C, Garcillán-Barcia MP, Francia MV, Rocha EP, de la Cruz F: Mobility of plasmids. Microbiol Mol Biol Rev 2010;74:434–452.
[PubMed]
21.
Smith MCA, Thomas CD: An accessory protein is required for relaxosome formation by small staphylococcal plasmids. J Bacteriol 2004;186:3363–3373.
[PubMed]
22.
van Zyl LJ, Deane SM, Rawlings DE: Analysis of the mobilization region of the broad-host-range IncQ-like plasmid pTC-F14 and its ability to interact with a related plasmid, pTF-FC2. J Bacteriol 2003;185:6104–6111.
[PubMed]
23.
Yamada Y, Yamada M, Nakazawa A: A ColE1-encoded gene directs entry exclusion of the plasmid. J Bacteriol 1995;177:6064–6068.
[PubMed]
24.
Zechner EL, de la Cruz F, Eisenbrandt R, Grahn AM, Koraimann G, Lanka E: Conjugative-DNA transfer processes; in Thomas CM (ed): The Horizontal Gene Pool: Bacterial Plasmids and Gene Spread. Amsterdam, Harwood Academic Publishers, 2000, pp 87–174.
© 2012 S. Karger AG, Basel
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
2012
You do not currently have access to this content.
