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RNA-structural mimicry in Escherichia coli ribosomal protein L4-dependent regulation of the S10 operon

Item Type:Article
Title:RNA-structural mimicry in Escherichia coli ribosomal protein L4-dependent regulation of the S10 operon
Creators Name:Stelzl, U. and Zengel, J.M. and Tovbina, M. and Walker, M. and Nierhaus, K.H. and Lindahl, L. and Patel, D.J.
Abstract:Ribosomal protein L4 regulates the 11-gene S10 operon in Escherichia coli by acting, in concert with transcription factor NusA, to cause premature transcription termination at a Rho-independent termination site in the leader sequence. This process presumably involves L4 interaction with the leader mRNA. Here, we report direct, specific, and independent binding of ribosomal protein L4 to the S10 mRNA leader in vitro. Most of the binding energy is contributed by a small hairpin structure within the leader region, but a 64-nucleotide sequence is required for the bona fide interaction. Binding to the S10 leader mRNA is competed by the 23 S rRNA L4 binding site. Although the secondary structures of the mRNA and rRNA binding sites appear different, phosphorothioate footprinting of the L4-RNA complexes reveals close structural similarity in three dimensions. Mutational analysis of the mRNA binding site is compatible with the structural model. In vitro binding of L4 induces structural changes of the S10 leader RNA, providing a first clue for how protein L4 may provoke transcription termination.
Keywords:5 Untranslated Regions, Amino Acid Sequence, Amino Acid Sequence Homology, Base Sequence, Binding Sites, Competitive Binding, Collodion, DNA Mutational Analysis, Drug Dose-Response Relationship, Enzymologic Gene Expression Regulation, Escherichia Coli, Genetic Transcription, Iodine, Messenger RNA, Molecular Models, Molecular Sequence Data, Nucleic Acid Conformation, Phylogeny, Protein Binding, Ribosomal 23S RNA, Ribosomal Proteins, Secondary Protein Structure
Source:Journal of Biological Chemistry
ISSN:0021-9258
Publisher:American Society for Biochemistry and Molecular Biology (U.S.A.)
Volume:278
Number:30
Page Range:28237-28245
Date:1 January 2003
Official Publication:https://doi.org/10.1074/jbc.M302651200
PubMed:View item in PubMed

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