ABSTRACT
The role of ribonucleases in the control of gene expression remains unknown in lactic acid bacteria. In the present work, we analysed the expression of the citP gene, which encodes the lactococcal citrate permease P, through the stability of the citQRP messenger in both Lactococcus lactis biovar diacetylactis (L. diacetylactis) and Escherichia coli. The chemical half-life for citQRP mRNA observed in L. diacetylactis wild-type strain was abnormally long for bacteria. It was even longer than that detected in E. coli RNase E or RNase III mutant strains. A model of processing and fate of RNA species containing citP gene is presented.
Subject(s)
Bacterial Proteins , Escherichia coli/enzymology , Lactococcus lactis/enzymology , Membrane Transport Proteins/genetics , Organic Anion Transporters , RNA, Bacterial/metabolism , RNA, Messenger/metabolism , Escherichia coli/genetics , Gene Expression Regulation, Enzymologic , Half-Life , Industrial Microbiology , Lactococcus lactis/genetics , Membrane Transport Proteins/metabolism , RNA, Bacterial/drug effects , RNA, Messenger/drug effects , Ribonucleases/pharmacologyABSTRACT
Citrate transport in Lactococcus lactis biovar diacetylactis (L. diacetylactis) is catalyzed by citrate permease P (CitP), which is encoded by the plasmidic citP gene. Two partial overlapping open reading frames citQ and citR are located upstream of citP. These two genes, together with citP, constitute the citQRPoperon. In this report it was shown that in L. diacetylactis and Escherichia coli, cit mRNA is subject to the same specific cleavages at a complex secondary structure which includes the central region of citQ and the 5'-end of citR. The role of ribonucleases in the fate of the cit mRNA processing was investigated in E. coli RNase mutant strains. The results obtained indicate that both endoribonucleases RNase E and RNase III are involved in the generation of mRNA processed species. RNase E is responsible for the major cleavages detected within citQ and upstream of citR, whereas RNase III cleaves citR within its ribosomal binding site. Preliminary results indicate the existence of a RNaselll-like enzyme in L. diacetylactis. Based on these results, a model for the role of cit mRNA processing in the expression of citP is presented.
Subject(s)
Bacterial Proteins/genetics , Carrier Proteins/genetics , Endoribonucleases/physiology , Escherichia coli Proteins , Lactococcus lactis/genetics , Operon , RNA Processing, Post-Transcriptional , RNA, Bacterial/metabolism , RNA, Messenger/metabolism , Base Sequence , Escherichia coli/enzymology , Molecular Sequence Data , Nucleic Acid Conformation , Ribonuclease III , SymportersABSTRACT
Citrate transport in Lactococcus lactis subsp. lactis biovar diacetylactis is catalyzed by citrate permease P (CitP), which is encoded by the plasmidic citP gene. We have shown previously that citP is included in the citQRP operon, which is mainly transcribed from the P1 promoter in L. lactis subsp. lactis biovar diacetylactis. furthermore, transcription of citQRP and citrate transport are not induced by the presence of citrate in the growth medium. In this work, we analyzed the influence of the extracellular pH on the expression of citP. The citrate transport system is induced by natural acidification of the medium during cell growth and by a shift to media buffered at acidic pHs. This inducible response to acid stress takes place at the transcriptional level and seems to be due to increased utilization of the P1 promoter. Increased transcription correlates with increased synthesis of CitP and results in higher citrate transport activity catalyzed by the cells. Finally, this acid stress response seems to provide L. lactis subsp. lactis biovar diacetylactis with a selective advantage resulting from cometabolism of glucose and citrate at low pHs.
