Open AccessLipase A gene transcription in P seudomonas alcaligenes is under control of RNA polymerase σ54 and response regulator LipR
Author(s) -
Krzeslak Joanna,
Papaioannou Evelina,
Merkerk Ronald,
Paal Krisztina A.,
Bischoff Rainer,
Cool Robbert H.,
Quax Wim J.
Publication year - 2012
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2012.02516.x
Subject(s) - response regulator , microbiology and biotechnology , enhancer , transcription (linguistics) , biology , rna polymerase , promoter , operon , gene , gene expression , transcriptional regulation , biochemistry , rna , mutant , linguistics , philosophy
Abstract Initial analysis has shown that the transcription of the P seudomonas alcaligenes lipA gene, which encodes an extracellular lipase, is governed by the Lip QR 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 ATP ase 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 A sp52 is the phosphorylated aspartate. This analysis exposes LipR as a prominent member of the growing family of bacterial enhancer‐binding proteins.