Structural and biophysical studies of transcriptional regulatory and anti-termination complexes
Lead Research Organisation:
MRC National Inst for Medical Research
Abstract
The work involves the application of biophysical and structural methods to investigate the mechanism of transcriptional regulation.
Technical Summary
Transcriptional anti-termination is a ubiquitous mechanism responsible for regulation and control of gene expression in eubacteria, archaebacteria, eukaryotic viruses and probably also in eukaryotes. The basic tenet of any antitermination mechanism is that the RNA polymerase (RNAP) becomes modified in such a way that it is able to readthrough termination signals allowing it to transcribe into a downstream gene, or further into an operon. The long term goal and objective of this work is to obtain detailed biochemical, biophysical and structural information concerning transcriptional anti-termination complexes in an attempt to provide insight into this important and ubiquitous mechanism of gene regulation. To do this we plan to undertake two main areas of study. The first will concentrate on the identification of components of anti-termination complexes using immunoprecipitation methods combined with mass spectroscopy. The second will focus on the detailed in vitro characterisation of the proteins and complexes involved in antitermination. This will combine structural data obtained from X-ray crystallographic, cryo-electron microscopy and magnetic resonance methods with kinetic and equilibrium binding data obtained from biophysical and biochemical measurements.||Initially, we will focus on the identification and characterisation of the components of antitermination complexes derived from E. coli and M. tuberculosis. However, this will be quickly extended to look at complexes from other organisms such as Mycobacterium leprae and the related but non-pathogenic Mycobacterium phlei and Mycobacterium smegmatis. As rRNA synthesis is intimately linked with bacterial growth rate we will be able to make a valuable comparison between the composition of antitermination complexes from slow growing pathogenic bacteria and the fast growing non-pathogenic varieties. We will also investigate sensor kinase directed antitermination mechanisms, initially concentrating on the proteins from the well-characterised E. coli and B. subtilis Bgl and Sac systems. However, we would also like to look at these systems in slow growing pathogenic mycobacteria, where the control of nutrient uptake is an important issue and sensor kinase directed antitermination mechanisms may well be involved.
People |
ORCID iD |
I Taylor (Principal Investigator) |
Publications

Beuth B
(2007)
Scaffold-independent analysis of RNA-protein interactions: the Nova-1 KH3-RNA complex.
in Journal of the American Chemical Society

Bishop KN
(2006)
Characterization of an amino-terminal dimerization domain from retroviral restriction factor Fv1.
in Journal of virology

Goldstone D
(2014)
Structural studies of postentry restriction factors reveal antiparallel dimers that enable avid binding to the HIV-1 capsid lattice
in Proceedings of the National Academy of Sciences

Goldstone D
(2013)
A Unique Spumavirus Gag N-terminal Domain with Functional Properties of Orthoretroviral Matrix and Capsid
in PLoS Pathogens

Goldstone DC
(2010)
Structural and functional analysis of prehistoric lentiviruses uncovers an ancient molecular interface.
in Cell host & microbe

Goldstone DC
(2011)
HIV-1 restriction factor SAMHD1 is a deoxynucleoside triphosphate triphosphohydrolase.
in Nature

Hilditch L
(2011)
Ordered assembly of murine leukemia virus capsid protein on lipid nanotubes directs specific binding by the restriction factor, Fv1.
in Proceedings of the National Academy of Sciences of the United States of America

Hollingworth D
(2006)
RNA polymerase II CTD phosphopeptides compete with RNA for the interaction with Pcf11.
in RNA (New York, N.Y.)

Ivins FJ
(2009)
NEMO oligomerization and its ubiquitin-binding properties.
in The Biochemical journal

Li J
(2008)
Chk2 oligomerization studied by phosphopeptide ligation: implications for regulation and phosphodependent interactions.
in The Journal of biological chemistry
Description | Structural studies of JSRV Gag proteins |
Organisation | University of Glasgow |
Department | School of Veterinary Medicine Glasgow |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | structural analysis of the JSRV gag proteins |
Collaborator Contribution | undertook complementary virological studies and together with the structural studies in my lab resulted in publication |
Impact | publication of paper in 2009 Pubmed ID 19007792 |
Start Year | 2007 |
Description | Structure and function of SAMHD1 |
Organisation | University of Manchester |
Department | School of Medicine Manchester |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | My team determined the structure and elucidated the mechanism of the HIV-1 restriction factor SAMHD1 |
Collaborator Contribution | identifying the SamHD1 gene and demonstrating it's role in Aicardi-Goutieres syndrome |
Impact | Structure of SAMHD1 determined findings published in Nature 2011 and Plos pathogens 2015 |
Start Year | 2016 |