The active form of the KorB protein encoded by the Streptomyces plasmid pIJ101 is a processed product that binds differentially to the two promoters it regulates

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Published inJournal of Bacteriology Vol. 175; no. 21; pp. 6996 - 7005
Main Authors Tai, J T, Cohen, S N
Format Journal Article
LanguageEnglish
Published Washington, DC American Society for Microbiology 01.11.1993
Subjects
Amino Acid Sequence
Bacterial Proteins - biosynthesis
Bacterial Proteins - isolation & purification
Bacterial Proteins - metabolism
Bacteriology
Bacteriophage T7 - genetics
Base Sequence
Binding Sites
Biological and medical sciences
Blotting, Western
Electrophoresis, Polyacrylamide Gel
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation, Bacterial
Genes, Bacterial
Genetics
Microbiology
Molecular Sequence Data
Molecular Weight
Plasmids
Promoter Regions, Genetic
Protein Binding
Proteins
Regulatory Sequences, Nucleic Acid
Repressor Proteins - biosynthesis
Repressor Proteins - isolation & purification
Repressor Proteins - metabolism
Restriction Mapping
Streptomyces - genetics
Streptomyces - metabolism
Streptomyces lividans
Streptomycetaceae
Molecular processing
DNA binding protein
Binding site
Structure function relationship
Genetic transfer
Actinomycetales
Posttranslational modification
Plasmid
Regulation(control)
Streptomyces lividans
Bacteria
Actinomycetes
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Abstract Article Usage Stats Services JB Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue JB About JB Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy JB RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0021-9193 Online ISSN: 1098-5530 Copyright © 2014 by the American Society for Microbiology.   For an alternate route to JB .asm.org, visit: JB       
AbstractList The partial purification and characterization of the KorB protein is reported, and some of its properties were investigated. The korB gene may have a role in coordinating the replication and intramycelial spread of plasmids during and/or following bacterial mating.
The korB gene of Streptomyces lividans plasmid pIJ101 is known to encode an autoregulated protein that also represses transcription of a gene, kilB, implicated in pIJ101 transfer and in spreading of the plasmid along mycelia of the recipient. Earlier work has indicated that the primary gene product of korB is a 10-kDa protein predicted from the gene sequence (D.S. Stein and S.N. Cohen, Mol. Gen. Genet. 222:337-344, 1990; S. Zamen H. Richards, and J. Ward, Nuleic Acids Res. 20:3693-3700, 1992). We report here that the 10-kDa KorB protein product is processed in vivo into a 6-kDa peptide that has a 20-fold-greater binding affinity for its operator-promoter target; in addition, the 6-kDa peptide binds differentially to the regulatory regions of the two genes it controls, showing 50-fold-greater affinity for the kilB sequence. While both the processed and unprocessed forms of KorB were observed in Escherichia coli following korB gene expression under control of the bacteriophage T7 promoter, only the 6-kDa peptide was found in S. lividans containing pIJ101, implying that this peptide is normally the biologically active form of KorB. The footprint resulting from KorB binding to the korB operator sequence overlaps the sti locus, which affects pIJ101 copy number and incompatibility as well as the size of zones of inhibited recipient cell growth ("pocks") that form around donor cells during mating. The observed ability of the korB gene product to interact with both sti sequences and the kilB promoter region suggests that it may have a role in coordinating the replication and intramycelial spread of plasmids during and/or following bacterial mating.
The korB gene of Streptomyces lividans plasmid pIJ101 is known to encode an autoregulated protein that also represses transcription of a gene, kilB, implicated in pIJ101 transfer and in spreading of the plasmid along mycelia of the recipient. Earlier work has indicated that the primary gene product of korB is a 10-kDa protein predicted from the gene sequence. We report here that the 10-kDa KorB protein product is processed in vivo into a 6-kDa peptide that has a 20-fold-greater binding affinity for its operator-promoter target; in addition, the 6-kDa peptide binds differentially to the regulatory regions of the two genes it controls, showing 50-fold-greater affinity for the kilB sequence. While both the processed and unprocessed forms of KorB were observed in Escherichia coli following korB gene expression under control of the bacteriophage T7 promoter, only the 6-kDa peptide was found in S. lividans containing pIJ101, implying that this peptide is normally the biologically active form of KorB. The footprint resulting from KorB binding to the korB operator sequence overlaps the sti locus, which affects pIJ101 copy number and incompatibility as well as the size of zones of inhibited recipient cell growth ("pocks") that form around donor cells during mating. The observed ability of the korB gene product to interact with both sti sequences and the kilB promoter region suggests that it may have a role in coordinating the replication and intramycelial spread of plasmids during and/or following bacterial mating.
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Author S N Cohen
J T Tai
AuthorAffiliation Department of Genetics, Stanford University School of Medicine, California 94305
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Issue 21
Keywords Streptomycetaceae
Molecular processing
DNA binding protein
Binding site
Structure function relationship
Genetic transfer
Actinomycetales
Posttranslational modification
Plasmid
Regulation(control)
Streptomyces lividans
Bacteria
Actinomycetes
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SSID ssj0014452
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Snippet Article Usage Stats Services JB Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley...
The korB gene of Streptomyces lividans plasmid pIJ101 is known to encode an autoregulated protein that also represses transcription of a gene, kilB, implicated...
The partial purification and characterization of the KorB protein is reported, and some of its properties were investigated. The korB gene may have a role in...
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StartPage 6996
SubjectTerms Amino Acid Sequence
Bacterial Proteins - biosynthesis
Bacterial Proteins - isolation & purification
Bacterial Proteins - metabolism
Bacteriology
Bacteriophage T7 - genetics
Base Sequence
Binding Sites
Biological and medical sciences
Blotting, Western
Electrophoresis, Polyacrylamide Gel
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation, Bacterial
Genes, Bacterial
Genetics
Microbiology
Molecular Sequence Data
Molecular Weight
Plasmids
Promoter Regions, Genetic
Protein Binding
Proteins
Regulatory Sequences, Nucleic Acid
Repressor Proteins - biosynthesis
Repressor Proteins - isolation & purification
Repressor Proteins - metabolism
Restriction Mapping
Streptomyces - genetics
Streptomyces - metabolism
Streptomyces lividans
Title The active form of the KorB protein encoded by the Streptomyces plasmid pIJ101 is a processed product that binds differentially to the two promoters it regulates
URI http://jb.asm.org/content/175/21/6996.abstract
https://www.ncbi.nlm.nih.gov/pubmed/8226643
https://www.proquest.com/docview/227085971
https://search.proquest.com/docview/16747731
https://search.proquest.com/docview/76052522
https://pubmed.ncbi.nlm.nih.gov/PMC206827
Volume 175
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