Accéder au contenu
Merck

A Novel Feedback Loop That Controls Bimodal Expression of Genetic Competence.

PLoS genetics (2015-06-26)
Pamela Gamba, Martijs J Jonker, Leendert W Hamoen
RÉSUMÉ

Gene expression can be highly heterogeneous in isogenic cell populations. An extreme type of heterogeneity is the so-called bistable or bimodal expression, whereby a cell can differentiate into two alternative expression states. Stochastic fluctuations of protein levels, also referred to as noise, provide the necessary source of heterogeneity that must be amplified by specific genetic circuits in order to obtain a bimodal response. A classical model of bimodal differentiation is the activation of genetic competence in Bacillus subtilis. The competence transcription factor ComK activates transcription of its own gene, and an intricate regulatory network controls the switch to competence and ensures its reversibility. However, it is noise in ComK expression that determines which cells activate the ComK autostimulatory loop and become competent for genetic transformation. Despite its important role in bimodal gene expression, noise remains difficult to investigate due to its inherent stochastic nature. We adapted an artificial autostimulatory loop that bypasses all known ComK regulators to screen for possible factors that affect noise. This led to the identification of a novel protein Kre (YkyB) that controls the bimodal regulation of ComK. Interestingly, Kre appears to modulate the induction of ComK by affecting the stability of comK mRNA. The protein influences the expression of many genes, however, Kre is only found in bacteria that contain a ComK homologue and, importantly, kre expression itself is downregulated by ComK. The evolutionary significance of this new feedback loop for the reduction of transcriptional noise in comK expression is discussed. Our findings show the importance of mRNA stability in bimodal regulation, a factor that requires more attention when studying and modelling this non-deterministic developmental mechanism.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
Phosphate de potassium monobasic, powder, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
Sigma-Aldrich
MES, low moisture content, ≥99% (titration)
Sigma-Aldrich
BIS-TRIS, ≥98.0% (titration)
Sigma-Aldrich
Rifampicine, ≥95% (HPLC), powder or crystals
Sigma-Aldrich
Acide éthylènediaminetétraacétique solution, 0.02% in DPBS (0.5 mM), sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Phosphate de potassium monobasic, for molecular biology, ≥98.0%
Sigma-Aldrich
Ethylenediaminetetraacetic acid, anhydrous, crystalline, BioReagent, suitable for cell culture
SAFC
BIS-TRIS
Sigma-Aldrich
Ethylenediaminetetraacetic acid, 99.995% trace metals basis
Sigma-Aldrich
Phosphate de potassium dibasic, anhydrous, for luminescence, for molecular biology, BioUltra, ≥99.0% (T)
Sigma-Aldrich
Potassium, chunks (in mineral oil), 98% trace metals basis
Sigma-Aldrich
Rifampicine, suitable for plant cell culture, BioReagent, ≥95% (HPLC), powder or crystals
Sigma-Aldrich
Potassium phosphate dibasic solution, 1.0 M
Sigma-Aldrich
Potassium hydride, 30 wt % dispersion in mineral oil
Sigma-Aldrich
Ethylenediaminetetraacetic acid, ACS reagent, 99.4-100.6%, powder
Sigma-Aldrich
BIS-TRIS, BioUltra, ≥99.0% (NT)
Sigma-Aldrich
BIS-TRIS, BioPerformance Certified, suitable for cell culture, suitable for insect cell culture, ≥98.0%
Sigma-Aldrich
BIS-TRIS, BioXtra, ≥98.0% (titration)
Sigma-Aldrich
Phosphate de potassium monobasic, BioUltra, for molecular biology, anhydrous, ≥99.5% (T)
Sigma-Aldrich
Phosphate de potassium monobasic, ReagentPlus®
Sigma-Aldrich
MES solution, BioUltra, for molecular biology, 0.5 M in H2O
Sigma-Aldrich
Phosphate de potassium monobasic, 99.99% trace metals basis
Sigma-Aldrich
Ethylenediaminetetraacetic acid, anhydrous, BioUltra, ≥99% (titration)
Sigma-Aldrich
Potassium hydride, in paraffin
Sigma-Aldrich
Phosphate de potassium dibasic, 99.95% trace metals basis
Sigma-Aldrich
Phosphate de potassium dibasic, reagent grade, ≥98.0%
SAFC
BIS-TRIS
Sigma-Aldrich
Ethylenediaminetetraacetic acid, purified grade, ≥98.5%, powder
Sigma-Aldrich
Potassium phosphate dibasic, meets USP testing specifications