An mRNA structure in bacteria that controls gene expression by binding lysine.

Riboswitches are metabolite-responsive genetic control elements that reside in the untranslated regions (UTRs) of certain messenger RNAs. Herein, we report that the 5'-UTR of the lysC gene of Bacillus subtilis carries a conserved RNA element that serves as a lysine-responsive riboswitch. The ligand-binding domain of the riboswitch binds to L-lysine with an apparent dissociation constant (KD) of approximately 1 micro M, and exhibits a high level of molecular discrimination against closely related analogs, including D-lysine and ornithine. Furthermore, we provide evidence that this widespread class of riboswitches serves as a target for the antimetabolite S-(2-aminoethyl)-L-cysteine (AEC). These findings add support to the hypotheses that direct sensing of metabolites by messenger RNAs is a fundamental form of genetic control and that riboswitches represent a new class of antimicrobial drug targets.

[1]  J. Szulmajster,et al.  Regulation of dihydrodipicolinate synthase and aspartate kinase in Bacillus subtilis , 1975, Journal of bacteriology.

[2]  Michael Hecker,et al.  Transcriptome and Proteome Analysis of Bacillus subtilis Gene Expression Modulated by Amino Acid Availability , 2002, Journal of bacteriology.

[3]  Evgeny Nudler,et al.  Sensing Small Molecules by Nascent RNA A Mechanism to Control Transcription in Bacteria , 2002, Cell.

[4]  S. Gottesman,et al.  Stealth regulation: biological circuits with small RNA switches. , 2002, Genes & development.

[5]  Y. Lu,et al.  Identification of aecA mutations in Bacillus subtilis as nucleotide substitutions in the untranslated leader region of the aspartokinase II operon. , 1991, Journal of general microbiology.

[6]  Ali Nahvi,et al.  An mRNA structure that controls gene expression by binding S-adenosylmethionine , 2003, Nature Structural Biology.

[7]  P. Zamecnik,et al.  Thiosine-resistant mutants of Escherichia coli K-12 with growth-medium-dependent lysl-tRNA synthetase activity. I. Isolation and physiological characterization. , 1972, Biochimica et biophysica acta.

[8]  C. Chan,et al.  Quantitative analysis of transcriptional pausing by Escherichia coli RNA polymerase: his leader pause site as paradigm. , 1996, Methods in enzymology.

[9]  Ali Nahvi,et al.  Genetic control by a metabolite binding mRNA. , 2002, Chemistry & biology.

[10]  R R Breaker,et al.  Relationship between internucleotide linkage geometry and the stability of RNA. , 1999, RNA.

[11]  V. Méjean,et al.  The leader sequence of the Escherichia coli lysC gene is involved in the regulation of LysC synthesis. , 1998, FEMS microbiology letters.

[12]  Jeffrey E. Barrick,et al.  Metabolite-binding RNA domains are present in the genes of eukaryotes. , 2003, RNA.

[13]  H. Liao,et al.  Analysis of the regulatory region of the lysC gene of Escherichia coli. , 1998, FEMS microbiology letters.

[14]  M. Flickinger,et al.  Cloning and nucleotide sequence of the gene coding for aspartokinase II from a thermophilic methylotrophic Bacillus sp , 1992, Applied and environmental microbiology.

[15]  Y. Lu,et al.  Fine-structure mapping of cis-acting control sites in the lysC operon of Bacillus subtilis. , 1992, FEMS microbiology letters.

[16]  R. Breaker,et al.  Immobilized RNA switches for the analysis of complex chemical and biological mixtures , 2001, Nature Biotechnology.

[17]  Ronald R. Breaker,et al.  Thiamine derivatives bind messenger RNAs directly to regulate bacterial gene expression , 2002, Nature.

[18]  A. Ellington,et al.  A (ribo) switch in the paradigms of genetic regulation , 2002, Nature Structural Biology.

[19]  E. Westhof,et al.  A common motif organizes the structure of multi-helix loops in 16 S and 23 S ribosomal RNAs. , 1998, Journal of molecular biology.

[20]  M. Ptashne,et al.  Genes and Signals , 2001 .

[21]  C. Yanofsky,et al.  Regulation by transcription attenuation in bacteria: how RNA provides instructions for transcription termination/antitermination decisions. , 2002, BioEssays : news and reviews in molecular, cellular and developmental biology.

[22]  R. Coccia,et al.  Aspartokinase III repression and lysine analogs utilization for protein synthesis. , 1990, Physiological chemistry and physics and medical NMR.

[23]  G. Bennett,et al.  Nucleotide sequence of the Escherichia coli cad operon: a system for neutralization of low extracellular pH , 1992, Journal of bacteriology.

[24]  R. Breaker,et al.  An mRNA structure that controls gene expression by binding FMN , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[25]  T. Henkin,et al.  Transcription termination control of the S box system: Direct measurement of S-adenosylmethionine by the leader RNA , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Jeffrey E. Barrick,et al.  Riboswitches Control Fundamental Biochemical Pathways in Bacillus subtilis and Other Bacteria , 2003, Cell.

[27]  B. Belitsky Biosynthesis of Amino Acids of the Glutamate and Aspartate Families, Alanine, and Polyamines , 2002 .

[28]  S. Kochhar,et al.  Lysine-induced premature transcription termination in the lysC operon of Bacillus subtilis. , 1996, Microbiology.

[29]  R. Losick,et al.  Bacillus Subtilis and Its Closest Relatives: From Genes to Cells , 2001 .

[30]  Vitaly Epshtein,et al.  The riboswitch-mediated control of sulfur metabolism in bacteria , 2003, Proceedings of the National Academy of Sciences of the United States of America.