Expression of bacteriocin LsbB is dependent on a transcription terminator

Gordana Uzelac; Marija Miljkovic; Jelena Lozo; Zorica Radulovic; Natasa Tosic; Milan Kojic

doi:10.1016/j.micres.2015.06.011

Expression of bacteriocin LsbB is dependent on a transcription terminator

Microbiol Res. 2015 Oct:179:45-53. doi: 10.1016/j.micres.2015.06.011. Epub 2015 Jul 16.

Authors

Gordana Uzelac¹, Marija Miljkovic², Jelena Lozo³, Zorica Radulovic⁴, Natasa Tosic⁵, Milan Kojic⁶

Affiliations

¹ Laboratory for Molecular Microbiology, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Serbia; Faculty of Agriculture, University of Belgrade, Serbia.
² Laboratory for Molecular Microbiology, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Serbia.
³ Laboratory for Molecular Microbiology, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Serbia; Faculty of Biology, University of Belgrade, Serbia.
⁴ Faculty of Agriculture, University of Belgrade, Serbia.
⁵ Laboratory for Molecular Biomedicine, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Serbia.
⁶ Laboratory for Molecular Microbiology, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Serbia. Electronic address: mkojic@imgge.bg.ac.rs.

PMID: 26411894
DOI: 10.1016/j.micres.2015.06.011

Abstract

The production of LsbB, leaderless class II bacteriocin, is encoded by genes (lsbB and lmrB) located on plasmid pMN5 in Lactococcus lactis BGMN1-5. Heterologous expression of the lsbB gene using the pAZIL vector (pAZIL-lsbB) in L. lactis subsp. cremoris MG7284 resulted in a significant reduction (more than 30 times) of bacteriocin LsbB expression. Subcloning and deletion experiments with plasmid pMN5 revealed that full expression of LsbB requires the presence of a complete transcription terminator located downstream of the lsbB gene. RNA stability analysis revealed that the presence of a transcription terminator increased the RNA stability by three times and the expression of LsbB by 30 times. The study of the influence of transcription terminator on the expression of other bacteriocin genes (lcnB, for lactococcin B production) indicated that this translational terminator likely functions in a lsbB-specific manner rather than in a general manner.

Keywords: Bacteriocin expression; LsbB baceriocin; Transcription terminator regulation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacteriocins / biosynthesis*
Bacteriocins / genetics
DNA, Bacterial*
Gene Expression Regulation, Bacterial
Gene Knockout Techniques
Genes, Bacterial*
Lactococcus lactis* / enzymology
Lactococcus lactis* / genetics
Plasmids
Promoter Regions, Genetic
Terminator Regions, Genetic
Transcription, Genetic

Substances

Bacteriocins
DNA, Bacterial
lactococcin B