Structure, function, and regulation of the bacterial transcription cycle

Information

  • Research Project
  • 10388954
  • ApplicationId
    10388954
  • Core Project Number
    R35GM118130
  • Full Project Number
    3R35GM118130-06S1
  • Serial Number
    118130
  • FOA Number
    PA-20-272
  • Sub Project Id
  • Project Start Date
    5/1/2016 - 8 years ago
  • Project End Date
    4/30/2026 - a year from now
  • Program Officer Name
    ADKINS, RONALD
  • Budget Start Date
    5/1/2021 - 3 years ago
  • Budget End Date
    4/30/2022 - 2 years ago
  • Fiscal Year
    2021
  • Support Year
    06
  • Suffix
    S1
  • Award Notice Date
    9/17/2021 - 2 years ago
Organizations

Structure, function, and regulation of the bacterial transcription cycle

Project Summary Transcription is the major control point of gene expression and RNA polymerase (RNAP), conserved from bacteria to man, is the central enzyme of transcription. Our long term goal is to understand the mechanism of transcription and its regulation. Determining three-dimensional structures of RNAP and its complexes with DNA, RNA, and regulatory factors, is an essential step. We focus on highly characterized prokaryotic RNAPs. The basic elements of the transcription cycle, initiation, elongation, and termination, were elucidated through study of prokaryotes. A detailed structural and functional understanding of the entire transcription cycle is essential to explain the fundamental control of gene expression and to target RNAP with small-molecule antibiotics. Advances in this understanding are stuck on the difficulty of visualizing transient intermediates that underlie the key transitions between stable states of the transcription cycle, and the difficulty of visualizing complex macromolecular assemblies involved in regulation, structural problems where X-ray crystallography has severe limitations. While the stable RNAP states around the transcription cycle (RNAP catalytic core, RNAP holoenzyme, RNAP holoenzyme open promoter complex, RNAP elongation complex) are relatively well characterized and understood, the transitions between the stable states are poorly understood. Major transitions include: Holoenzyme + promoter DNA è? open promoter complex (initiation) Open promoter complex è? elongation complex (promoter escape, ? dissociation) Elongation complex è? core RNAP + DNA + completed RNA transcript (termination) Each of these transitions are characterized by unstable, transient intermediates that are extremely challenging for structural biology. At every stage of the transcription cycle, RNAP function is modulated by interactions with extrinsic regulatory factors. Assembling and crystallizing transcription complexes containing extrinsic regulators also presents challenges for structural biology. Due to recent advances, cryo-electron microscopy (cryo-EM) now offers a route to structural and mechanistic characterization of these intermediates and large assemblies. We will use cryo-electron microscopy, in combination with X-ray crystallography and other approaches, to exploit this opportunity and provide a complete characterization of the bacterial transcription cycle.

IC Name
NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
  • Activity
    R35
  • Administering IC
    GM
  • Application Type
    3
  • Direct Cost Amount
    53560
  • Indirect Cost Amount
  • Total Cost
    53560
  • Sub Project Total Cost
  • ARRA Funded
    False
  • CFDA Code
    859
  • Ed Inst. Type
    GRADUATE SCHOOLS
  • Funding ICs
    NIGMS:53560\
  • Funding Mechanism
    Non-SBIR/STTR RPGs
  • Study Section
    ZRG1
  • Study Section Name
    Special Emphasis Panel
  • Organization Name
    ROCKEFELLER UNIVERSITY
  • Organization Department
    PHYSIOLOGY
  • Organization DUNS
    071037113
  • Organization City
    NEW YORK
  • Organization State
    NY
  • Organization Country
    UNITED STATES
  • Organization Zip Code
    100656399
  • Organization District
    UNITED STATES