Lipase A gene transcription in Pseudomonas alcaligenes is under control of RNA polymerase σ54 and response regulator LipR.
Initial analysis has shown that the transcription of the Pseudomonas alcaligenes lipA gene, which encodes an extracellular lipase, is governed by the LipQR two-component system consisting of sensor kinase LipQ and DNA-binding regulator LipR. This study further analyzes lipA gene expression and demonstrates that the RNA polymerase σ54 is involved in the transcription. Purified LipR has an ATPase activity that is stimulated by the presence of lipA promoter DNA. Surface plasmon resonance measurements with purified and in vitro phosphorylated LipR reveal that phosphorylation of LipR is required for specific binding to the upstream activating sequence of the lipA promoter. Furthermore, mass spectrometric analysis combined with mutagenesis demonstrates that Asp52 is the phosphorylated aspartate. This analysis exposes LipR as a prominent member of the growing family of bacterial enhancer-binding proteins.
Department of Pharmaceutical Biology, University Center of Pharmacy, Groningen University Institute for Drug Exploration, Groningen, The Netherlands.
Joanna
J
Krzeslak
Evelina
E
Papaioannou
Ronald
R
van Merkerk
Krisztina A
KA
Paal
Rainer
R
Bischoff
Robbert H
RH
Cool
Wim J
WJ
Quax
D000251:Adenosine Triphosphatases / Q000378:metabolism; D001224:Aspartic Acid / Q000378:metabolism; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D008049:Lipase / Q000096:biosynthesis / Q000235:genetics*; D009154:Mutation; D010766:Phosphorylation; D044202:Pseudomonas alcaligenes / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D051877:RNA Polymerase Sigma 54 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000235:genetics*
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D014157:Transcription Factors; D001224:Aspartic Acid; D051877:RNA Polymerase Sigma 54; C113518:LipA protein, Pseudomonas; D008049:Lipase; D000251:Adenosine Triphosphatases