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The bacteriophage T4 transcription activator MotA interacts with the far-C-terminal region of the sigma70 subunit of Escherichia coli RNA polymerase. 2002

Suchira Pande, and Anna Makela, and Simon L Dove, and Bryce E Nickels, and Ann Hochschild, and Deborah M Hinton
Laboratory of Molecular and Cellular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892-0830, USA.

Transcription from bacteriophage T4 middle promoters uses Escherichia coli RNA polymerase together with the T4 transcriptional activator MotA and the T4 coactivator AsiA. AsiA binds tightly within the C-terminal portion of the sigma70 subunit of RNA polymerase, while MotA binds to the 9-bp MotA box motif, which is centered at -30, and also interacts with sigma70. We show here that the N-terminal half of MotA (MotA(NTD)), which is thought to include the activation domain, interacts with the C-terminal region of sigma70 in an E. coli two-hybrid assay. Replacement of the C-terminal 17 residues of sigma70 with comparable sigma38 residues abolishes the interaction with MotA(NTD) in this assay, as does the introduction of the amino acid substitution R608C. Furthermore, in vitro transcription experiments indicate that a polymerase reconstituted with a sigma70 that lacks C-terminal amino acids 604 to 613 or 608 to 613 is defective for MotA-dependent activation. We also show that a proteolyzed fragment of MotA that contains the C-terminal half (MotA(CTD)) binds DNA with a K(D(app)) that is similar to that of full-length MotA. Our results support a model for MotA-dependent activation in which protein-protein contact between DNA-bound MotA and the far-C-terminal region of sigma70 helps to substitute functionally for an interaction between sigma70 and a promoter -35 element.

UI MeSH Term Description Entries
D004247 DNA A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine). DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D004268 DNA-Binding Proteins Proteins which bind to DNA. The family includes proteins which bind to both double- and single-stranded DNA and also includes specific DNA binding proteins in serum which can be used as markers for malignant diseases. DNA Helix Destabilizing Proteins,DNA-Binding Protein,Single-Stranded DNA Binding Proteins,DNA Binding Protein,DNA Single-Stranded Binding Protein,SS DNA BP,Single-Stranded DNA-Binding Protein,Binding Protein, DNA,DNA Binding Proteins,DNA Single Stranded Binding Protein,DNA-Binding Protein, Single-Stranded,Protein, DNA-Binding,Single Stranded DNA Binding Protein,Single Stranded DNA Binding Proteins
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D012321 DNA-Directed RNA Polymerases Enzymes that catalyze DNA template-directed extension of the 3'-end of an RNA strand one nucleotide at a time. They can initiate a chain de novo. In eukaryotes, three forms of the enzyme have been distinguished on the basis of sensitivity to alpha-amanitin, and the type of RNA synthesized. (From Enzyme Nomenclature, 1992). DNA-Dependent RNA Polymerases,RNA Polymerases,Transcriptases,DNA-Directed RNA Polymerase,RNA Polymerase,Transcriptase,DNA Dependent RNA Polymerases,DNA Directed RNA Polymerase,DNA Directed RNA Polymerases,Polymerase, DNA-Directed RNA,Polymerase, RNA,Polymerases, DNA-Dependent RNA,Polymerases, DNA-Directed RNA,Polymerases, RNA,RNA Polymerase, DNA-Directed,RNA Polymerases, DNA-Dependent,RNA Polymerases, DNA-Directed
D012808 Sigma Factor A protein which is a subunit of RNA polymerase. It effects initiation of specific RNA chains from DNA. Sigma Element,Sigma Initiation Factor,Sigma Subunit,Minor Sigma Factor,RNA Polymerase Sigma Factor H,Factor, Sigma,Factor, Sigma Initiation,Initiation Factor, Sigma,Sigma Factor, Minor,Subunit, Sigma
D014157 Transcription Factors Endogenous substances, usually proteins, which are effective in the initiation, stimulation, or termination of the genetic transcription process. Transcription Factor,Factor, Transcription,Factors, Transcription
D014158 Transcription, Genetic The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION. Genetic Transcription
D014764 Viral Proteins Proteins found in any species of virus. Gene Products, Viral,Viral Gene Products,Viral Gene Proteins,Viral Protein,Protein, Viral,Proteins, Viral
D020798 Two-Hybrid System Techniques Screening techniques first developed in yeast to identify genes encoding interacting proteins. Variations are used to evaluate interplay between proteins and other molecules. Two-hybrid techniques refer to analysis for protein-protein interactions, one-hybrid for DNA-protein interactions, three-hybrid interactions for RNA-protein interactions or ligand-based interactions. Reverse n-hybrid techniques refer to analysis for mutations or other small molecules that dissociate known interactions. One-Hybrid System Techniques,Reverse One-Hybrid System Techniques,Reverse Two-Hybrid System Techniques,Three-Hybrid System Techniques,Yeast Two-Hybrid Assay,Yeast Two-Hybrid System Techniques,One-Hybrid System Technics,Reverse Three-Hybrid System Techniques,Three-Hybrid System Technics,Tri-Hybrid System Techniques,Two-Hybrid Assay,Two-Hybrid Method,Two-Hybrid System Technics,Yeast One-Hybrid System Techniques,Yeast Three-Hybrid Assay,Yeast Three-Hybrid System,Yeast Three-Hybrid System Techniques,Yeast Two-Hybrid System,n-Hybrid System Techniques,Assay, Two-Hybrid,Assay, Yeast Three-Hybrid,Assay, Yeast Two-Hybrid,Assays, Two-Hybrid,Assays, Yeast Three-Hybrid,Assays, Yeast Two-Hybrid,Method, Two-Hybrid,Methods, Two-Hybrid,One Hybrid System Technics,One Hybrid System Techniques,One-Hybrid System Technic,One-Hybrid System Technique,Reverse One Hybrid System Techniques,Reverse Three Hybrid System Techniques,Reverse Two Hybrid System Techniques,System Technique, n-Hybrid,System Techniques, n-Hybrid,System, Yeast Three-Hybrid,System, Yeast Two-Hybrid,Systems, Yeast Three-Hybrid,Systems, Yeast Two-Hybrid,Technic, One-Hybrid System,Technic, Three-Hybrid System,Technic, Two-Hybrid System,Technics, One-Hybrid System,Technics, Three-Hybrid System,Technics, Two-Hybrid System,Technique, One-Hybrid System,Technique, Three-Hybrid System,Technique, Tri-Hybrid System,Technique, Two-Hybrid System,Technique, n-Hybrid System,Techniques, One-Hybrid System,Techniques, Three-Hybrid System,Techniques, Tri-Hybrid System,Techniques, Two-Hybrid System,Techniques, n-Hybrid System,Three Hybrid System Technics,Three Hybrid System Techniques,Three-Hybrid Assay, Yeast,Three-Hybrid Assays, Yeast,Three-Hybrid System Technic,Three-Hybrid System Technique,Three-Hybrid System, Yeast,Three-Hybrid Systems, Yeast,Tri Hybrid System Techniques,Tri-Hybrid System Technique,Two Hybrid Assay,Two Hybrid Method,Two Hybrid System Technics,Two Hybrid System Techniques,Two-Hybrid Assay, Yeast,Two-Hybrid Assays,Two-Hybrid Assays, Yeast,Two-Hybrid Methods,Two-Hybrid System Technic,Two-Hybrid System Technique,Two-Hybrid System, Yeast,Two-Hybrid Systems, Yeast,Yeast One Hybrid System Techniques,Yeast Three Hybrid Assay,Yeast Three Hybrid System,Yeast Three Hybrid System Techniques,Yeast Three-Hybrid Assays,Yeast Three-Hybrid Systems,Yeast Two Hybrid Assay,Yeast Two Hybrid System,Yeast Two Hybrid System Techniques,Yeast Two-Hybrid Assays,Yeast Two-Hybrid Systems,n Hybrid System Techniques,n-Hybrid System Technique

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