RRC ID 3322
著者 Liu Y, Ichige A, Kobayashi I.
タイトル Regulation of the EcoRI restriction-modification system: Identification of ecoRIM gene promoters and their upstream negative regulators in the ecoRIR gene.
ジャーナル Gene
Abstract Type II restriction-modification (R-M) systems are composed of linked restriction endonuclease and modification methyltransferase genes and serve as barriers to horizontal gene transfer even though they are mobile in themselves. Their products kill host bacterial cells that have lost the R-M genes, a process that helps to maintain the frequency of the R-M systems in the viable cell population. Their establishment and maintenance in a bacterial host are expected to involve fine regulation of their gene expression. In the present study, we analyzed transcription of the modification gene and its regulation within the EcoRI R-M system. Northern blotting revealed that the downstream ecoRIM gene is transcribed as a monocistronic mRNA and as part of a larger bicistronic mRNA together with the upstream ecoRIR gene. Primer extension, RNase protection, and mutational analysis using lacZ gene fusions identified two overlapping promoters for ecoRIM gene transcription within the ecoRIR gene. Further mutational analysis revealed that two upstream AT-rich elements within the ecoRIR gene, "AATAAA" and "ATTATAAATATA," function as negative regulators of these promoters. Simultaneous substitution of these two elements resulted in a four-fold increase in beta-galactosidase activity and a five-fold increase in transcript levels as measured by RNase protection assay. RNA measurements of the ecoRIM transcript suggested that these elements decreased ecoRIM expression by interfering with transcription initiation of the ecoRIM promoters. Possible roles for these ecoRIM promoters and their negative regulators in the EcoRI R-M system are discussed.
巻・号 400(1-2)
ページ 140-9
公開日 2007-10-1
DOI 10.1016/j.gene.2007.06.006
PII S0378-1119(07)00311-3
PMID 17618069
MeSH Base Sequence DNA Restriction-Modification Enzymes / genetics* Deoxyribonuclease EcoRI / genetics Escherichia coli / genetics* Gene Expression Regulation, Bacterial* Gene Expression Regulation, Enzymologic* Genes, Bacterial Molecular Sequence Data Promoter Regions, Genetic* Regulatory Sequences, Nucleic Acid Site-Specific DNA-Methyltransferase (Adenine-Specific) / genetics Transcription Initiation Site
IF 2.984
引用数 10
WOS 分野 GENETICS & HEREDITY
リソース情報
原核生物(大腸菌) pAB2001