Gene expression and pathogenicity in mycobacteria

Lead Research Organisation: MRC National Inst for Medical Research

Abstract

Tuberculosis (TB) is the leading cause of death worldwide due to an infectious disease, a situation exacerbated by its deadly synergy with AIDS. Annually approximately 2 million people die from TB. Even in the UK and other western countries there is a continuing significant increase in the disease in recent years. The situation is not helped by the limited effectiveness, at least in some part of the world, of the widely used vaccine BCG. Moreover there is a great threat from bacteria that are resistant to treatment with conventional anti-TB drugs, resulting in a disease that is essentially non-treatable unless new antibiotics are discovered. Thus strategies for developing new approaches to the treatment and prevention of TB are a high public health priority. Research at the National Institute for Medical Research is aimed at trying to gain a better understanding of the way in which the bacterium which causes TB is able to infect people and how that process might be interrupted. Using molecular genetics the researchers are identifying some of the key molecules produced by the bacterium and the infected cells, which enable the infection to either progress or be eliminated. These molecules are then being used to screen for novel anti-TB drugs in collaboration with MRC-Technology.

Technical Summary

Mycobacteria are a group of bacteria that include some of the most important causes of human infectious disease. Different mycobacterial species are the causes of tuberculosis (TB), the leading cause of death worldwide due to an infectious disease, and leprosy, a disease that results in deformity and disability in millions of people. The objective of this work is to understand the mechanisms which underlie the pathogenicity of the causative agent of human TB, Mycobacterium tuberculosis and to search for new therapeutic agents against it. In this project we are attempting to identify key molecules of M. tuberculosis which enable it to survive and grow inside macrophages, the host cells which are invaded by the bacteria in these diseases. We can also monitor the ways in which the macrophages respond to this invasion by the bacteria. By understanding this interaction we hope to develop new approaches for treating and/or preventing these diseases. This work involves the use of genetic approaches to identify molecules which are important for regulating gene expression and for growth and survival of M. tuberculosis inside macrophages. By introducing mutations into bacteria we can ask whether such changes reduce the ability of the bacteria to cause disease; if so, then the genes which we have mutated can be characterised to give us a better understanding of their function and thus to provide important information about how the bacteria cause TB. We are also able to analyse changes in gene expression that occur within macrophages following infection in order to identify key molecules which are produced by the host and which may influence the outcome of the infection. The changes in gene expression, in both bacteria and host cells, are monitored using whole genome DNA microarrays, either a spotted microarray for M. tuberculosis, or an Affymetrix microarray for host (mouse or human) cells. Having identified candidate genes in M. tuberculosis, deletions of them are derived using homologous recombination techniques, and the effect of the deletion is investigated both by infecting macrophages in vitro and by infecting mice. Identification of the key molecules involved in determining the outcome of infection is a starting point for identifying potential new drug targets. We can produce recombinant protein molecules and develop assays as the basis for screens for new anti-TB drugs. These high throughput screens are performed in collaboration with MRC-Technology. Since the emergence of multiple drug-resistant strains of M. tuberculosis and limitations in the effectiveness of the existing vaccine BCG represent an enormous public health challenge, such approaches are extremely important for the future control of TB and related diseases.

Publications

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Crack JC (2011) Mechanistic insight into the nitrosylation of the [4Fe-4S] cluster of WhiB-like proteins. in Journal of the American Chemical Society

 
Description International Joint Projects 2008/R2-2009
Amount £12,000 (GBP)
Organisation The Royal Society 
Sector Academic/University
Country United Kingdom
Start 02/2009 
End 02/2011
 
Description Joint Project Grant-2007
Amount £95,254 (GBP)
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 09/2007 
End 09/2009
 
Description MRCT Development Gap Funding
Amount £107,126 (GBP)
Organisation MRC-Technology 
Sector Academic/University
Country United Kingdom
Start 09/2006 
End 09/2008
 
Title EspR mutant. 
Description Deletion of espR (Rv3849) gene of Mycobacterium tuberculosis. 
Type Of Material Cell line 
Year Produced 2011 
Provided To Others? Yes  
Impact PMID: 22155774. PMID: 22389481. PMID: 22722242. 
URL http://europepmc.org/abstract/MED/22155774
 
Title PknF mutant. 
Description Deletion of pknF gene in Mycobacterium tuberculosis. 
Type Of Material Cell line 
Year Produced 2012 
Provided To Others? Yes  
Impact PMID: 21622570. PMID: 23915284. 
URL http://europepmc.org/abstract/MED/21622570
 
Title Rv1747 mutant. 
Description Deletion of Rv1747 gene in Mycobacterium tuberculosis H37Rv. 
Type Of Material Cell line 
Provided To Others? No  
Impact PMID: 16040957. PMID: 21622570. 
URL http://europepmc.org/abstract/MED/16040957
 
Title Rv3616c mutant 
Description Deletion of Rv3616c (espA) gene in Mycobacterium tuberculosis. 
Type Of Material Cell line 
Year Produced 2012 
Provided To Others? Yes  
Impact None as yet. 
 
Title Rv3676 mutant. 
Description Deletion of Rv3676 gene in Mycobacterium tuberculosis H37Rv. 
Type Of Material Cell line 
Year Produced 2009 
Provided To Others? Yes  
Impact PMID: 15882420. PMID: 20028978. PMID: 21902733. 
URL http://europepmc.org/abstract/MED/15882420
 
Title WhiB1 mutant 
Description Deletion of whiB1 (Rv3219) gene of Mycobacterium tuberculosis. 
Type Of Material Cell line 
Year Produced 2012 
Provided To Others? Yes  
Impact PMID: 20929442. 
URL http://europepmc.org/abstract/MED/20929442
 
Description Evaluation of inhibitors of Mycobacterium tuberculosis protein kinases. 
Organisation MRC-Technology
Department MRCT Centre for Therapeutics Discovery (CTD)
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution In vitro and in vivo experiments to evaluate lead compounds, discussion and analysis of results.
Collaborator Contribution Screening of compound libraries, discussion and analysis of results.Intellectual and experimental.
Impact MRC-Technology Development Gap Fund award - 2006. PMID:21482481. PMID: 22469702.
 
Description Evaluation of inhibitors of Mycobacterium tuberculosis protein kinases. 
Organisation Medical Research Council (MRC)
Department MRC National Institute for Medical Research (NIMR)
Country United Kingdom 
Sector Public 
PI Contribution In vitro and in vivo experiments to evaluate lead compounds, discussion and analysis of results.
Collaborator Contribution Screening of compound libraries, discussion and analysis of results.Intellectual and experimental.
Impact MRC-Technology Development Gap Fund award - 2006. PMID:21482481. PMID: 22469702.
 
Description Mechanism of action of the WhiB1 protein. 
Organisation Medical Research Council (MRC)
Department MRC National Institute for Medical Research (NIMR)
Country United Kingdom 
Sector Public 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Intellectual and experimental.Intellectual and experimental.Intellectual and experimental.
Impact PMID: 20929442. PMID: 21182249. PMID: 22464736. PMID: 22792304.
Start Year 2009
 
Description Mechanism of action of the WhiB1 protein. 
Organisation University of East Anglia
Department School of Biological Sciences UEA
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Intellectual and experimental.Intellectual and experimental.Intellectual and experimental.
Impact PMID: 20929442. PMID: 21182249. PMID: 22464736. PMID: 22792304.
Start Year 2009
 
Description Mechanism of action of the WhiB1 protein. 
Organisation University of Leicester
Department Department of Infection, Immunity and Inflammation
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Intellectual and experimental.Intellectual and experimental.Intellectual and experimental.
Impact PMID: 20929442. PMID: 21182249. PMID: 22464736. PMID: 22792304.
Start Year 2009
 
Description Mechanism of action of the WhiB1 protein. 
Organisation University of Sheffield
Department Department of Molecular Biology and Biotechnology
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Intellectual and experimental.Intellectual and experimental.Intellectual and experimental.
Impact PMID: 20929442. PMID: 21182249. PMID: 22464736. PMID: 22792304.
Start Year 2009
 
Description Mode of action of the EspR transcriptional regulator in Mycobacterium tuberculosis. 
Organisation University of Sheffield
Department Department of Infection, Immunity and Cardiovascular Disease
Country United Kingdom 
Sector Academic/University 
PI Contribution Intellectual and experimental contributions.
Collaborator Contribution Intellectual and experimental contributions.
Impact PMID: 22389481. PMID: 22722242
 
Description Mycobacterium tuberculosis strains in a UK South Indian community. 
Organisation Birmingham Heartlands Hospital
Country United Kingdom 
Sector Hospitals 
PI Contribution Designing and perfroming experiments and analysis of results.
Collaborator Contribution Provision of DNA from clinical strains of Mycobacterium tuberculosis.Provision of DNA from clinical strains of Mycobacterium tuberculosis.Provision of DNA microarrays.Provision of DNA from clinical strains of Mycobacterium tuberculosis.
Impact PMID: 17719277.
 
Description Mycobacterium tuberculosis strains in a UK South Indian community. 
Organisation St George's University of London
Department Department of Cellular and Molecular Medicine
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and perfroming experiments and analysis of results.
Collaborator Contribution Provision of DNA from clinical strains of Mycobacterium tuberculosis.Provision of DNA from clinical strains of Mycobacterium tuberculosis.Provision of DNA microarrays.Provision of DNA from clinical strains of Mycobacterium tuberculosis.
Impact PMID: 17719277.
 
Description Mycobacterium tuberculosis strains in a UK South Indian community. 
Organisation University of Birmingham
Department College of Medical and Dental Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and perfroming experiments and analysis of results.
Collaborator Contribution Provision of DNA from clinical strains of Mycobacterium tuberculosis.Provision of DNA from clinical strains of Mycobacterium tuberculosis.Provision of DNA microarrays.Provision of DNA from clinical strains of Mycobacterium tuberculosis.
Impact PMID: 17719277.
 
Description Mycobacterium tuberculosis strains in a UK South Indian community. 
Organisation University of Leeds
Department School of Biochemistry and Microbiology Leeds
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and perfroming experiments and analysis of results.
Collaborator Contribution Provision of DNA from clinical strains of Mycobacterium tuberculosis.Provision of DNA from clinical strains of Mycobacterium tuberculosis.Provision of DNA microarrays.Provision of DNA from clinical strains of Mycobacterium tuberculosis.
Impact PMID: 17719277.
 
Description New antituberculosis agents amongst known drugs. 
Organisation MRC-Technology
Department MRCT Centre for Therapeutics Discovery (CTD)
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution Designing and performing of experiments and analysis of results.
Collaborator Contribution Designing of experiments and provision of compound library.
Impact PMID: 19699151.
Start Year 2006
 
Description Regulation of protein kinase activity in Mycobacterium tuberculosis. 
Organisation Medical Research Council (MRC)
Department MRC National Institute for Medical Research (NIMR)
Country United Kingdom 
Sector Public 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Designing and performing experiments and analysis of results.Designing and performing experiments and analysis of results.Designing and performing experiments and analysis of results.
Impact PMID: 16040957. PMID: 19318624. PMID: 21134638. PMID: 21482481. PMID: 21622570.
 
Description Regulation of protein kinase activity in Mycobacterium tuberculosis. 
Organisation University of Leicester
Department Department of Infection, Immunity and Inflammation
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Designing and performing experiments and analysis of results.Designing and performing experiments and analysis of results.Designing and performing experiments and analysis of results.
Impact PMID: 16040957. PMID: 19318624. PMID: 21134638. PMID: 21482481. PMID: 21622570.
 
Description Regulation of protein kinase activity in Mycobacterium tuberculosis. 
Organisation University of Montpellier
Country France 
Sector Academic/University 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Designing and performing experiments and analysis of results.Designing and performing experiments and analysis of results.Designing and performing experiments and analysis of results.
Impact PMID: 16040957. PMID: 19318624. PMID: 21134638. PMID: 21482481. PMID: 21622570.
 
Description Regulation of the ESX-1 secretion system in Mycobacterium tuberculosis 
Organisation Swiss Federal Institute of Technology in Lausanne (EPFL)
Department Global Health Institute
Country Switzerland 
Sector Academic/University 
PI Contribution Intellectual and experimental contributions. Provision of mutant bacteria.
Collaborator Contribution Intellectual and experimental contributions.
Impact PMID: 22155774
Start Year 2011
 
Description Role of cAMP in mycobacteria. 
Organisation Indian Institute of Science, Bangalore
Department Department of Molecular Reproduction, Development and Genetics (MRDG)
Country India 
Sector Academic/University 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Intellectual discussions, experimental and provision of resources.
Impact Royal Society International Travel Award - 2009.
Start Year 2006
 
Description Use of DNA microarrays to analyse genome expression in the macrophage. 
Organisation Imperial College London
Department MRC Centre for Molecular Bacteriology and Infection
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Designing and performing experiments and analysis of results.Designing and performing experimenst and analysis of results.
Impact PMID: 16782391. Yang, M; Krishnan, N; Coade, S; Buxton, RS; Hubank, M; Young, DB and Tascon, RE (2009) Activation by CpG oligodeoxynucleotide protects bone marrow derived dendritic cells from apoptosis: A transcriptomic and bio-informatic study. The Open Immunology Journal 2, 67-78
 
Description Use of DNA microarrays to analyse genome expression in the macrophage. 
Organisation Medical Research Council (MRC)
Department MRC National Institute for Medical Research (NIMR)
Country United Kingdom 
Sector Public 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Designing and performing experiments and analysis of results.Designing and performing experimenst and analysis of results.
Impact PMID: 16782391. Yang, M; Krishnan, N; Coade, S; Buxton, RS; Hubank, M; Young, DB and Tascon, RE (2009) Activation by CpG oligodeoxynucleotide protects bone marrow derived dendritic cells from apoptosis: A transcriptomic and bio-informatic study. The Open Immunology Journal 2, 67-78
 
Description Use of chromatin immunoprecipitation to study transcription factor distribution on the Mycobacterium tuberculosis chromosome. 
Organisation Medical Research Council (MRC)
Department MRC National Institute for Medical Research (NIMR)
Country United Kingdom 
Sector Public 
PI Contribution Intellectual and experimental contributions.
Collaborator Contribution Intellectual and experimental contributions.
Impact PMID: 22528497.
Start Year 2008
 
Description cAMP receptor protein of Mycobacterium bovis BCG. 
Organisation Medical Research Council (MRC)
Department MRC National Institute for Medical Research (NIMR)
Country United Kingdom 
Sector Public 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Discussion of experiment design and results.Discussion of experiment design and results.Analysis of protein structures.
Impact PMID: 18332206.
 
Description cAMP receptor protein of Mycobacterium bovis BCG. 
Organisation University of Birmingham
Department College of Life and Environmental Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Discussion of experiment design and results.Discussion of experiment design and results.Analysis of protein structures.
Impact PMID: 18332206.
 
Description cAMP receptor protein of Mycobacterium bovis BCG. 
Organisation University of Birmingham
Department College of Medical and Dental Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Discussion of experiment design and results.Discussion of experiment design and results.Analysis of protein structures.
Impact PMID: 18332206.
 
Description cAMP receptor protein of Mycobacterium tuberculosis. 
Organisation St George's University of London
Department Department of Cellular and Molecular Medicine
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Intellectual and experimental contributions.Provision of DNA microarrays.
Impact PMID: 15882420. Wellcome joint Project Grant (078731/Z/05/Z) with University of Sheffield - 2007. PMID:20028978. PMID:20929442.
 
Description cAMP receptor protein of Mycobacterium tuberculosis. 
Organisation University of Leicester
Department Department of Infection, Immunity and Inflammation
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Intellectual and experimental contributions.Provision of DNA microarrays.
Impact PMID: 15882420. Wellcome joint Project Grant (078731/Z/05/Z) with University of Sheffield - 2007. PMID:20028978. PMID:20929442.
 
Description cAMP receptor protein of Mycobacterium tuberculosis. 
Organisation University of Sheffield
Country United Kingdom 
Sector Academic/University 
PI Contribution Designing and performing experiments and analysis of results.
Collaborator Contribution Intellectual and experimental contributions.Provision of DNA microarrays.
Impact PMID: 15882420. Wellcome joint Project Grant (078731/Z/05/Z) with University of Sheffield - 2007. PMID:20028978. PMID:20929442.
 
Description Schools visits - NIMR 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach Local
Primary Audience Schools
Results and Impact Talks given to primary school students.

NA
Year(s) Of Engagement Activity 2008,2009,2010