A5013500340- RNA and protein synthesis mechanisms
- Bacterial Genetics and Biotechnology
- Bacteriophages and microbial interactions
- Microbial Natural Products and Biosynthesis
- Genomics and Phylogenetic Studies
- RNA Research and Splicing
- Carbohydrate Chemistry and Synthesis
- DNA and Nucleic Acid Chemistry
- Plant-Microbe Interactions and Immunity
- RNA modifications and cancer
- Molecular Biology Techniques and Applications
- Animal Nutrition and Physiology
- Enzyme-mediated dye degradation
- Cancer therapeutics and mechanisms
- Glycosylation and Glycoproteins Research
- Microbial Metabolism and Applications
- Advanced Proteomics Techniques and Applications
- Entomopathogenic Microorganisms in Pest Control
- Animal Behavior and Welfare Studies
- Protein purification and stability
- Inhalation and Respiratory Drug Delivery
- Salmonella and Campylobacter epidemiology
- DNA Repair Mechanisms
- Effects of Environmental Stressors on Livestock
- Marine Sponges and Natural Products
University of Leeds
2011-2024
Institute of Structural and Molecular Biology
2002-2022
University of Sheffield
2014
University of Cambridge
2003-2005
Bangor University
2005
Daresbury Laboratory
2003-2005
University of Oxford
2005
Institute of Molecular Biology, Academia Sinica
1994-1998
Vienna Biocenter
1998
Stanford University
1993-1996
Abstract Ribonuclease E has a central role in Escherichia coli mRNA decay and is dependent on functional product of the rne (also called ams or hmp1) gene. We investigated requirements for RNase cleavage by introducing random mutations into decanucleotide region at 5' end pACYC184 RNA I studying effects these position rne-dependent vivo E-mediated cutting vitro. find that precise point can be altered specifically reproducibly sequence changes cleaved and, therefore, not determined distance...
Abstract Escherichia coli endoribonuclease E has a major influence on gene expression. It is essential for the maturation of ribosomal and transfer RNA as well rapid degradation messenger RNA. The latter ensures that translation closely follows programming at level transcription. Recently, one hallmarks RNase E, i.e. its ability to bind via 5′-monophosphorylated end, was shown be unnecessary initial cleavage some polycistronic tRNA precursors. Here we show using RNA-seq analyses...
Thernegene ofEscherichia coliencodes a 118 kDa protein that has ribonuclease E (RNase E) activity and binds RNA. A functionalrnegene product is essential for cell viability the processing and/or decay of variety RNA species, including 9 S RNA, mRNA RNAI, antisense regulator ColE1-type plasmid replication. By testing ability different segments Rne to catalyze cleavage bind we found N-terminal half (residues 1 498) contains catalytic function sufficient site-specific oligoribonucleotides...
Summary The Streptomyces produce a plethora of secondary metabolites including antibiotics and undergo complex developmental cycle. As means establishing the pathways that regulate metabolite production by this important bacterial genus, model species coelicolor its relatives have been subject several genetic screens. However, despite identification characterization numerous genes affect antibiotic production, there is still no overall understanding network integrates various environmental...
Escherichia coli RNase E, an essential single-stranded specific endoribonuclease, is required for both ribosomal RNA processing and the rapid degradation of mRNA. The availability complete sequences a number bacterial genomes prompted us to assess evolutionarily conservation E. We show here that sequence N-terminal endoribonucleolytic domain E conserved in Synechocystis sp. other bacteria. Furthermore, we demonstrate homologue binds substrates cleaves them at same position as enzyme. Taken...
RNase E, an endoribonuclease encoded by theEscherichia coli ~j m j h ~p l locus, cleaves RNA I, antisense regulator of the replication ColEl type plas-mid~, in a single-stranded region near its 6' end.The m-3071 mutation prolongs I half-life cells cultured at elevated temperature and imparts sensitivity on E isolated from mutant strain.Here we report effects specific sequence changes introduced site-directed mutagenesis location ribonucleolytic cleavage 5' end pBR322 congenic r n e + E....
Two temperature-sensitive mutations, ams-1 and rne-3071, in the ams (altered mRNA stability) gene have been used extensively to investigate processing decay of RNA Escherichia coli. We sequenced these alleles found that mutations are separated by only 6 nucleotides cause conservative amino acid substitutions next a possible nucleotide-binding site within N-terminal domain Ams protein. Computer analysis revealed region altered has extensive sequence similarity predicted product from mre...
The availability of nutrients is a major determinant for the timing morphogenesis and antibiotic production in soil-dwelling bacterium Streptomyces coelicolor. Here we show that N-acetylglucosamine transport, first step an important nutrient signalling cascade, mediated by NagE2 permease phosphotransferase system, activity this linked to nutritional control development production. serves as high-affinity transporter (K(m) 2.6 microM). complex was reconstituted with individually purified...
The best characterized pathway for the initiation of mRNA degradation in Escherichia coli involves removal 5′-terminal pyrophosphate to generate a monophosphate group that stimulates endonucleolytic cleavage by RNase E. We show here however, using well-characterized oligonucleotide substrates and transcripts, E can cleave certain RNAs rapidly without requiring 5′-monophosphorylated end. Moreover, minimum substrate requirement this mode cleavage, which be categorized as 'direct' or 'internal'...
Summary S treptomyces coelicolor is a model for studying bacteria renowned as the foremost source of natural products used clinically. Post‐genomic studies have revealed complex patterns gene expression and links to growth, morphological development individual genes. However, underlying regulation remains largely obscure, but undoubtedly involves steps after transcription initiation. Here we identify sites involved in RNA processing degradation well within nucleotide‐resolution map...
Summary The sequence of a 2657 bp DNA fragment containing the coding and regulatory regions oxytetracycline (OTC)‐resistance gene, otrA , from OTC producer Streptomyces rimosus was determined. predicted amino acid OtrA had extensive identity with tetracycline‐resistance genes other bacteria which mediate resistance via non‐covalent ribosomal modification. N ‐terminal domain extremely high GTP‐binding sites elongation factors, such as EF‐G EF‐Tu, suggesting that binding hydrolysis GTP is...
RNase E is an essential endoribonuclease that plays a central role in the processing and degradation of RNA Escherichia coli other bacteria. Most endoribonucleases have been shown to act distributively; however, Feng et al. [(2002) Proc. Natl. Acad. Sci. U.S.A. 99, 14746−14751] recently found acts via scanning mechanism. A structural explanation for processivity provided here, with our finding conserved catalytic domain E. forms homotetramer. Nondissociating nanoflow-electrospray mass...
The intricate regulation of the Escherichia coli rpoS gene, which encodes stationary phase sigma-factor σ S , includes translational activation by noncoding RNA DsrA. We observed that stability mRNA, and concomitantly concentration were significantly higher in an RNase III-deficient mutant. As no decay intermediates corresponding to vitro mapped III cleavage site leader could be detected vivo, initial appears decisive for rapid inactivation mRNA. In contrast, we show base-pairing DsrA with...
Abstract The RNase E family is renowned for being central to the processing and decay of all types RNA in many species bacteria, as well providing first examples endonucleases that can recognize 5′-monophosphorylated ends thereby increasing efficiency cleavage. However, there evidence some transcripts be cleaved efficiently by Escherichia coli via direct entry, i.e. absence recognition a end. Here, we provide biochemical entry transfer (tRNA) E. coli, one core functions E, show it mediated...
Summary The RNase E/G family of endoribonucleases has a central role in RNA degradation and processing. Previous work shown that their cleavage substrates vitro can be stimulated by the presence 5′ monophosphate group. It not however, established importance this activation for any natural processing or decay pathway vivo. Here we provide Escherichia coli G first evidence sensing is required vivo normal rapid functional mRNAs; moreover, show that, contrast to previous study, enhance affinity...
Abstract The Escherichia coli endoribonuclease RNase E is central to the processing and degradation of all types RNA as such a pleotropic regulator gene expression. It essential for growth was one first examples an endonuclease that can recognise 5′-monophosphorylated ends thereby increasing efficiency many cleavages. Homologues be found in bacterial families including important pathogens, but no homologues have been identified humans or animals. represents potential target development new...
Summary The control of secondary production in streptomycetes involves the funneling environmental and physiological signals to cluster‐situated (transcriptional) regulators ( CSRs ) biosynthetic genes. For some systems, binding products CSR has been shown provide negative feedback. Here we show for lidamycin C ‐1027), a clinically relevant antitumor agent, by S treptomyces globisporus that feedback can extend point higher regulatory cascade. We DNA ‐binding activity . orthologue AtrA ,...