Author: Uyen Nguyen
Publisher:
ISBN:
Category :
Languages : en
Pages :
Book Description
RNA polymerase (RNAP) is an essential enzyme that is responsible for transcription of genes in all organisms. A protein called HelD (YvgS) has been recently identified as a transcription factor of RNAP in Gram-positive Bacillus subtilis, which enhances transcription by increasing RNAP recycling. Based on sequence homology, HelD belongs to the Superfamily 1 (SF1) DNA helicase. However, the mechanism of HelD-dependent transcription activation is limited due to lack of the structures of HelD and also the RNAP and HelD complex. The goal of this project is to obtain a high-resolution three-dimensional structure of HelD in complex with B. subtilis RNAP to elucidate the structural basis of RNAP recycling by the DNA helicase HelD. Two techniques have been used toward the structure determinations in this project: one is X-ray crystallography to determine the structure of HelD, and single-particle cryo-electron microscopy (cryo-EM) to determine the structure of HelD-RNAP complex. Truncations of HelD were constructed for protein expression to enhance protein crystallibility. DNA binding and unwinding function were assayed with HelD using electrophoretic mobility shift assay to ensure functional HelD was purified for downstream structural determination experiments. Overall, this project provides a framework for purification protocol of HelD. In future experiments, structure determination of HelD and HelD- RNAP complex would enhance our knowledge of bacterial transcription.
Characterization of HelD, a Novel RNA Polymerase Binding Protein in Bacillus Subtilis
Author: Uyen Nguyen
Publisher:
ISBN:
Category :
Languages : en
Pages :
Book Description
RNA polymerase (RNAP) is an essential enzyme that is responsible for transcription of genes in all organisms. A protein called HelD (YvgS) has been recently identified as a transcription factor of RNAP in Gram-positive Bacillus subtilis, which enhances transcription by increasing RNAP recycling. Based on sequence homology, HelD belongs to the Superfamily 1 (SF1) DNA helicase. However, the mechanism of HelD-dependent transcription activation is limited due to lack of the structures of HelD and also the RNAP and HelD complex. The goal of this project is to obtain a high-resolution three-dimensional structure of HelD in complex with B. subtilis RNAP to elucidate the structural basis of RNAP recycling by the DNA helicase HelD. Two techniques have been used toward the structure determinations in this project: one is X-ray crystallography to determine the structure of HelD, and single-particle cryo-electron microscopy (cryo-EM) to determine the structure of HelD-RNAP complex. Truncations of HelD were constructed for protein expression to enhance protein crystallibility. DNA binding and unwinding function were assayed with HelD using electrophoretic mobility shift assay to ensure functional HelD was purified for downstream structural determination experiments. Overall, this project provides a framework for purification protocol of HelD. In future experiments, structure determination of HelD and HelD- RNAP complex would enhance our knowledge of bacterial transcription.
Publisher:
ISBN:
Category :
Languages : en
Pages :
Book Description
RNA polymerase (RNAP) is an essential enzyme that is responsible for transcription of genes in all organisms. A protein called HelD (YvgS) has been recently identified as a transcription factor of RNAP in Gram-positive Bacillus subtilis, which enhances transcription by increasing RNAP recycling. Based on sequence homology, HelD belongs to the Superfamily 1 (SF1) DNA helicase. However, the mechanism of HelD-dependent transcription activation is limited due to lack of the structures of HelD and also the RNAP and HelD complex. The goal of this project is to obtain a high-resolution three-dimensional structure of HelD in complex with B. subtilis RNAP to elucidate the structural basis of RNAP recycling by the DNA helicase HelD. Two techniques have been used toward the structure determinations in this project: one is X-ray crystallography to determine the structure of HelD, and single-particle cryo-electron microscopy (cryo-EM) to determine the structure of HelD-RNAP complex. Truncations of HelD were constructed for protein expression to enhance protein crystallibility. DNA binding and unwinding function were assayed with HelD using electrophoretic mobility shift assay to ensure functional HelD was purified for downstream structural determination experiments. Overall, this project provides a framework for purification protocol of HelD. In future experiments, structure determination of HelD and HelD- RNAP complex would enhance our knowledge of bacterial transcription.
Characterization of Promoters and Genes Controlled by Bacillus Subtilis Sigma-28 RNA Polymerase
Author: Victoria Lyn Singer
Publisher:
ISBN:
Category :
Languages : en
Pages : 470
Book Description
Publisher:
ISBN:
Category :
Languages : en
Pages : 470
Book Description
RNA Polymerase-binding Proteins from Sporulating Bacillus Subtilis
Author: Arno Lee Greenleaf
Publisher:
ISBN:
Category :
Languages : en
Pages :
Book Description
Publisher:
ISBN:
Category :
Languages : en
Pages :
Book Description
The Isolation and Characterization of the Bacillus Subtilis Gene Encoding the Major Sigma Factor of RNA Polymerase
Author: Michael Alan Gitt
Publisher:
ISBN:
Category :
Languages : en
Pages : 336
Book Description
Publisher:
ISBN:
Category :
Languages : en
Pages : 336
Book Description
Genetics of the Protein Secretion Machinery of Bacillus Subtilis with Characterization of a Novel Component, PrsA Lipoprotein
Author: Vesa Kontinen
Publisher:
ISBN: 9789514772092
Category :
Languages : en
Pages : 65
Book Description
Publisher:
ISBN: 9789514772092
Category :
Languages : en
Pages : 65
Book Description
Genetics and Biotechnology of Bacilli
Author: Mark M. Zukowski
Publisher: Elsevier
ISBN: 0323150624
Category : Nature
Languages : en
Pages : 432
Book Description
Genetics and Biotechnology of Bacilli, Volume 3 covers the proceedings of the Fifth International Conference on Genetics and Biotechnology of Bacilli, held on July 9-12, 1989 at the Asilomar Conference Center, Pacific Grove, California. It summarizes the remarkable progress made in the genetics and biotechnology fields of Bacilli. It is organized into four parts, encompassing 43 chapters, which focus on gene regulation and structure, enzyme structure, Bacillus thuringiensis toxins, and stationary phase gene regulation. Part I covers topics related to gene regulation and structure of Bacilli, such as control of gene expression, mutation, genetic organization, DNA sequence analysis, and identification of transcript units. It also discusses gene replication in Bacillus subtilis plasmids, levanase operon of B. subtilis, and characterization of global regulon in B. subtilis. The next part of this book focuses on the structure of various enzymes found in B. subtilis, including alpha amylases, subtilisin, alkaline phosphatase, and levansucrase. Part III discusses the generation of functional B. thuringiensis toxin hybrid genes, regulation of crystal protein gene promoters, toxicity of B. thuringiensis delta-endotoxin, and insecticidal activity of chimeric protoxins. The concluding part covers the aspects of signal transduction, regulation of differential gene expression during B. subtilis sporulation, and gene cloning and deletion for extracellular proteases of B. subtilis. It also discusses genetic and biochemical aspects of protein phosphorylation; properties of B. subtilis spores; control of stationary phase gene expression; and the novel regulatory gene, senS, of B. subtilis. This book is a valuable source of information for microbiologists, research biologists, and Bacilli enthusiasts.
Publisher: Elsevier
ISBN: 0323150624
Category : Nature
Languages : en
Pages : 432
Book Description
Genetics and Biotechnology of Bacilli, Volume 3 covers the proceedings of the Fifth International Conference on Genetics and Biotechnology of Bacilli, held on July 9-12, 1989 at the Asilomar Conference Center, Pacific Grove, California. It summarizes the remarkable progress made in the genetics and biotechnology fields of Bacilli. It is organized into four parts, encompassing 43 chapters, which focus on gene regulation and structure, enzyme structure, Bacillus thuringiensis toxins, and stationary phase gene regulation. Part I covers topics related to gene regulation and structure of Bacilli, such as control of gene expression, mutation, genetic organization, DNA sequence analysis, and identification of transcript units. It also discusses gene replication in Bacillus subtilis plasmids, levanase operon of B. subtilis, and characterization of global regulon in B. subtilis. The next part of this book focuses on the structure of various enzymes found in B. subtilis, including alpha amylases, subtilisin, alkaline phosphatase, and levansucrase. Part III discusses the generation of functional B. thuringiensis toxin hybrid genes, regulation of crystal protein gene promoters, toxicity of B. thuringiensis delta-endotoxin, and insecticidal activity of chimeric protoxins. The concluding part covers the aspects of signal transduction, regulation of differential gene expression during B. subtilis sporulation, and gene cloning and deletion for extracellular proteases of B. subtilis. It also discusses genetic and biochemical aspects of protein phosphorylation; properties of B. subtilis spores; control of stationary phase gene expression; and the novel regulatory gene, senS, of B. subtilis. This book is a valuable source of information for microbiologists, research biologists, and Bacilli enthusiasts.
RNA-protein Interaction
Author: Sai Lung Su
Publisher:
ISBN:
Category :
Languages : en
Pages : 262
Book Description
Publisher:
ISBN:
Category :
Languages : en
Pages : 262
Book Description
Revealing the Interactions of RNA Polymerase II Binding Proteins
Author: Roland Becker
Publisher:
ISBN:
Category :
Languages : en
Pages : 123
Book Description
Publisher:
ISBN:
Category :
Languages : en
Pages : 123
Book Description
An RNA Polymerase from Bacillus Subtilis Bearing a Novel Sigma Subunit
Author: Janey Lee Wiggs
Publisher:
ISBN:
Category :
Languages : en
Pages : 332
Book Description
Publisher:
ISBN:
Category :
Languages : en
Pages : 332
Book Description
Structural and Functional Studies of Bacillus Subtilis RNA Polymerase Sigma-A Factor
Author: Ban-Yang Chang
Publisher:
ISBN:
Category :
Languages : en
Pages : 354
Book Description
Publisher:
ISBN:
Category :
Languages : en
Pages : 354
Book Description