A novel, inducible, eukaryotic gene expression system based on the quorum-sensing transcription factor TraR.
Bacteria adapt their pattern of gene expression in response to a variety of external cues, including fluctuations in population density. This type of bacterial cell-to-cell communication is referred to as quorum-sensing. Quorum-sensing systems are present in many bacterial species and constitute a large collection of ligands and cognate receptors. The availability of such diversity offers interesting opportunities for biotechnological exploitation. We describe here the transformation of the quorum-sensing system of Agrobacterium tumefaciens into a transcription regulatory system that works in mammalian cells. The A. tumefaciens TraR protein was fused to the eukaryotic activation domain of NF-kappaB p65, generating a novel chimaeric transcriptional activator that stimulates gene transcription in different human cell lines from a minimal promoter containing the TraR DNA recognition sequence in the presence of the Agrobacterium quorum-sensing signal molecule N-(3-oxo-octanoyl)homoserine lactone (3-oxo-C(8)-HSL). The basal level of transcription was low in the absence of 3-oxo-C(8)-HSL, and gene expression was stimulated up to 1,000-fold at a saturating concentration of 3-oxo-C(8)-HSL.
Neddermann
Petra
P
Istituto di Ricerche di Biologia Molecolare P. Angeletti, Via Pontina Km 30.600, 00040 Pomezia/Roma, Italy. Petra_Neddermann@merck.com
Gargioli
Cesare
C
Muraglia
Ester
E
Sambucini
Sonia
S
Bonelli
Fabio
F
De Francesco
Raffaele
R
Cortese
Riccardo
R
D001426:Bacterial Proteins / Q000502:physiology*; D004247:DNA / Q000502:physiology*; D015870:Gene Expression / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D014158:Transcription, Genetic / Q000502:physiology
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; C080512:TraR protein, Agrobacterium tumefaciens; D004247:DNA