Authors

Pradervand, Nicolas Sulser, Sandra Delavat, François Miyazaki, Ryo Lamas, Iker van der Meer, Jan Roelof

Affiliations

Department of Fundamental Microbiology, University of Lausanne, Lausanne, Switzerland.

Application Area

Optogenetics

Time-Lapse Microscopy

Abstract

The integrative and conjugative element ICEclc is a mobile genetic element in Pseudomonas knackmussii B13, and an experimental model for a widely distributed group of elements in Proteobacteria. ICEclc is transferred from specialized transfer competent cells, which arise at a frequency of 3-5% in a population at stationary phase. Very little is known about the different factors that control the transfer frequency of this ICE family. Here we report the discovery of a three-gene operon encoded by ICEclc, which exerts global control on transfer initiation. The operon consists of three consecutive regulatory genes, encoding a TetR-type repressor MfsR, a MarR-type regulator and a LysR-type activator TciR. We show that MfsR autoregulates expression of the operon, whereas TciR is a global activator of ICEclc gene expression, but no clear role was yet found for MarR. Deletion of mfsR increases expression of tciR and marR, causing the proportion of transfer competent cells to reach almost 100% and transfer frequencies to approach 1 per donor. mfsR deletion also caused a two orders of magnitude loss in population viability, individual cell growth arrest and loss of ICEclc. This indicates that autoregulation is an important feature maintaining ICE transfer but avoiding fitness loss. Bioinformatic analysis showed that the mfsR-marR-tciR operon is unique for ICEclc and a few highly related ICE, whereas tciR orthologues occur more widely in a large variety of suspected ICE among Proteobacteria.

Extract

… “Inverted Microscope Eclipse Ti-E, equipped with a Perfect Focus System (PFS), pE-100 CoolLED and a”…

Product Associated Features

pE-100: A range of compact, simple-to-use, single wavelength illumination systems for screening fluorescence.

Diascopic Technique

Phase Contrast

Live Cell Issues

-

Product Type

pE-100

Journal

PloS one

Year of Publication

2014

Country of Publication

Switzerland