Copper resistance in salmonella
Lead Research Organisation:
University of Manchester
Department Name: Life Sciences
Abstract
Food-borne Salmonella infections cause serious medical and veterinary problems worldwide. In the developed world, salmonellosis continues to be one of the most common forms of food-poisoning and mainly occurs due to the consumption of contaminated animal products. Salmonella food poisoning represents a serious health risk and is usually characterised by the acute onset of fever, abdominal pain, diarrhea, nausea and occasionally vomiting but in some cases the disease can become more severe and life-threatening. Following oral ingestion of Salmonella, disease is started by the bacteria being able to survive within the intestinal tract and invade intestinal cells. The ability of the bacteria to reside in host immune cells (macrophages) is important for their spread to different organs and systemic disease. Survival within the different locations of the host requires that Salmonella can sense and adapt to changing environmental conditions such as varying metal levels. One such metal is copper. Copper is required by Salmonella for several key enzymes but is also extremely toxic. Our preliminary studies have now indicated that resistance to copper-stress plays a vital role during Salmonella infections. This project aims to build upon these studies and provide a detailed characterisation of the mechanisms of copper-resistance in this pathogen and their roles during infection. Furthermore, we will test the hypothesis that copper transporters in host cells contribute to their ability to kill bacteria by supplying copper to generate highly toxic reactive oxygen species. Understanding the environmental challenges that occur in the infected host and the mechanisms which protect Salmonella from copper stress may help in developing novel therapeutics to combat disease. Furthermore, there is a growing use of copper as a means of controlling bacterial growth in agriculture and the food industry. Clearly this has implications for the development of more resistant bacteria. Hence, a thorough understanding of the copper-resistance systems in Salmonella and their role in virulence is vital.
Technical Summary
This programme of work will test the hypothesis that the copper-resistance systems in Salmonella enterica serovar Typhimurium play an important role in pathogenicity. Obtaining copper for loading onto copper-dependent enzymes is a vital requirement for S. Typhimurium during infection. However, copper is extremely toxic and indeed is now thought to constitute a crucial component of the bacterial killing mechanisms within a host. S. Typhimurium must therefore possess systems that ensure a supply of copper to copper-requiring proteins whilst avoiding copper-toxicity. Our preliminary studies have shown that the cue and gol systems in S. Typhimurium both have roles in providing protection from copper-stress. We now intend to perform a detailed characterisation of these systems and examine the roles of the cue and gol proteins in copper-sensing, -transport and -trafficking under different stress conditions. It is anticipated that the possession of these two systems provides S. Typhimurium with the ability to adapt to excess cytoplasmic copper under different surplus copper conditions encountered at different locations within a host. This will be investigated. We will also test the hypothesis that mammalian copper transporters contribute to the anti-bacterial activity of macrophages by supplying copper for the Fenton reaction and hence the generation of deadly hydroxyl radicals. With the widespread use of copper to control microbial growth in agricultural practices and the food industry, a thorough investigation of the copper-resistance systems in S. Typhimurium and their role in virulence is both timely and necessary. It is anticipated that an understanding of the roles of the copper-resistance systems in S. Typhimurium infection could enhance the development of new drugs and control strategies.
Publications
Jennifer Cavet (Principle Investigator)
(2010)
Atypical copper homeostasis in Salmonella Typhimurium
Jennifer Cavet (Author)
(2010)
Atypical copper homeostasis in Salmonella typhimurium
Osman D
(2010)
Bacterial metal-sensing proteins exemplified by ArsR-SmtB family repressors.
in Natural product reports
Kimber RL
(2020)
Biomineralization of Cu2S Nanoparticles by Geobacter sulfurreducens.
in Applied and environmental microbiology
Cavet JS
(2014)
Copper as a magic bullet for targeted microbial killing.
in Chemistry & biology
Jennifer Cavet (Author)
(2012)
Copper homeostasis and Salmonella pathogenicity
Osman D
(2010)
Copper homeostasis in Salmonella is atypical and copper-CueP is a major periplasmic metal complex.
in The Journal of biological chemistry
Wang Y
(2010)
Elucidation of the functional metal binding profile of a Cd(II)/Pb(II) sensor CmtR(Sc) from Streptomyces coelicolor.
in Biochemistry
Ladomersky E
(2017)
Host and Pathogen Copper-Transporting P-Type ATPases Function Antagonistically during Salmonella Infection.
in Infection and immunity
Osman D
(2011)
Metal sensing in Salmonella: implications for pathogenesis.
in Advances in microbial physiology
Osman D
(2013)
The copper supply pathway to a Salmonella Cu,Zn-superoxide dismutase (SodCII) involves P(1B)-type ATPase copper efflux and periplasmic CueP.
in Molecular microbiology
Corbett D
(2012)
Two zinc uptake systems contribute to the full virulence of Listeria monocytogenes during growth in vitro and in vivo.
in Infection and immunity
Description | Salmonella is one of the most common causes of food-borne disease in man and represents a serious health risk. Disease mainly occurs due to the consumption of contaminated animal products and is usually characterised by the acute onset of fever, abdominal pain, diarrhoea, nausea and occasionally vomiting, but in some cases the disease can become more severe and life-threatening. The ability of Salmonella to cause disease is associated with its ability to survive and replicate inside the antimicrobial compartments, known as phagosomes, of host immune cells (macrophages). In this study we established that high copper levels are a feature of macrophage phagosomes and in response to bacterial infection macrophages increase production of their copper-transporters to concentrate copper within these compartments and exploit copper-toxicity in their anti-microbial defences. We demonstrated that Salmonella possesses two related copper-exporting proteins, CopA and GolT, and both have a role in copper resistance. Importantly, copper-resistance due to copper-export by CopA or GolT, is required for pathogen survival in macrophage phagosomes. In addition to a role in copper-resistance, we also demonstrated that both CopA and GolT are required in the copper-supply pathway for periplasmic Cu,Zn-superoxide dismutase (SOD). This is the first evidence of a copper-trafficking pathway for the activation of a periplasmic Cu,Zn-SOD. We identified a novel protein CueP as an abundant periplasmic copper-binding protein also required in the copper supply pathway for Salmonella Cu,Zn-SOD activation. Copper is directly transferred from Cu-CueP to pre-folded Zn-SOD to generate the active enzyme, implicating CueP as a copper-chaperone. |
Exploitation Route | The discovery that copper contributes to innate immune defences and that copper-resistance contributes to the ability of microbes to cause infections is being exploited in the development of copper-related antimicrobials. We have identified novel copper-homeostatic proteins in Salmonella that represent targets for antimicrobial drug-development. The copper-supply pathway that we have uncovered in Salmonella represents a paradigm system and knowledge of this system may be applied to improving small molecule, bioenergy industrial biotechnology and potentially bioprocessing applications. |
Sectors | Agriculture Food and Drink Manufacturing including Industrial Biotechology Pharmaceuticals and Medical Biotechnology |
Description | Public engagement activities with findings of this research: Used as an example to support the A-level syllabus via an article in the 'Biological Sciences Review' magazine for sixth formers (2014) Presented in a lecture to year 12 school pupils - Discover Life Sciences (2016). |
First Year Of Impact | 2014 |
Sector | Education |
Impact Types | Societal |
Description | Advanced Life Sciences Research Technology Initiative |
Amount | £409,319 (GBP) |
Funding ID | BB/L014823/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 01/2014 |
End | 03/2014 |
Description | Engineering Microbial Metal Recovery (EMMR) |
Amount | £844,637 (GBP) |
Funding ID | BB/W01467X/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 01/2022 |
End | 01/2024 |
Description | Food-borne Listeria infections: Zinc homeostasis at the host-pathogen interface |
Amount | £483,000 (GBP) |
Funding ID | BB/K016881/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 01/2014 |
End | 01/2017 |
Description | Metal-sensing in Salmonella: A model for targeting a network that differentiates metals |
Amount | £759,000 (GBP) |
Funding ID | BB/J017787/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 09/2012 |
End | 10/2017 |
Description | Molecular engineering of high activity multifunctional biometallic catalysts for clickable chemistries |
Amount | £713,334 (GBP) |
Funding ID | BB/R010412/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 03/2018 |
End | 03/2021 |
Description | Collaboration Leicester |
Organisation | University of Leicester |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Research collaboration leading to publications and future funding |
Collaborator Contribution | Research collaboration leading to publications and future funding |
Impact | Publications in research journals (Corbett et al., 2011 & 2012) and future funding (BB/K016881/1) |
Start Year | 2010 |
Description | International collaboration |
Organisation | Indiana University |
Country | United States |
Sector | Academic/University |
PI Contribution | Research collaboration leading to publication |
Collaborator Contribution | Research collaboration leading to publication |
Impact | Publication in research journal (Wang et al., 2010) |
Start Year | 2009 |
Description | Article for Biological Sciences Review |
Form Of Engagement Activity | A magazine, newsletter or online publication |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | Article for Biological Sciences Review (2014) Salmonella: A bacterial pathogen that invades our gut. Aimed at 6th formers to support A level syllabus and has resulted in requests for visits/placements from 6th formers. None so far |
Year(s) Of Engagement Activity | 2014 |
Description | British Science Week 2013 |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | NOWGEN DNA stand at Science Extravaganza for British Science Week (a 10 day event with engaging activities for people of all ages) - the activity was at Manchester Museum and hosted as a public event |
Year(s) Of Engagement Activity | 2012 |
Description | Copper Homeostasis and Salmonella Pathogenesis |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Type Of Presentation | poster presentation |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Invited speaker Invited speaker, Gordon Research Conference (GRC), Cell Biology of Metals, Rhode Island, USA. no actual impacts realised to date |
Year(s) Of Engagement Activity | 2011 |
Description | FLS open day |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Public/other audiences |
Results and Impact | Managing an interactive 'Exploring DNA Stand' at a faculty of life sciences (University of Manchester) community open day - aimed at engaging with the local community, The presentation sparked questions and discussions. |
Year(s) Of Engagement Activity | 2013 |
Description | Lecture to year 12 school pupils |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Schools |
Results and Impact | Research findings were presented to ~20 year 12 school pupils who visited the research organisation |
Year(s) Of Engagement Activity | 2014 |
Description | NorthWest Microbiology 2011 |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Type Of Presentation | paper presentation |
Geographic Reach | Regional |
Primary Audience | Postgraduate students |
Results and Impact | Speaker at North West Microbiology meeting aimed at young researchers - talk sparked questions and possible collaborations no actual impacts realised to date |
Year(s) Of Engagement Activity | 2011 |
Description | NorthWest Microbiology 2012 |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Postgraduate students |
Results and Impact | Presentations at North West Microbiology meeting aimed at young researchers - presentations sparked questions and possible collaborations |
Year(s) Of Engagement Activity | 2012 |
Description | Presentation at the 10th International Copper Meeting (2016) |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Poster presentation by researcher and postgraduate student at the 10th International Copper Meeting (Sorrento, Naples, Italy) |
Year(s) Of Engagement Activity | 2016 |
Description | Supervision of a Nuffield Research Placement |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Schools |
Results and Impact | Engagement of school 6th former in primary research - influenced career plans After this placements there have been requests to host future Nuffield placements |
Year(s) Of Engagement Activity | 2012 |