Prof C Baldock, Dr A Roseman, Dr T Jowitt
No more applications being accepted
Competition Funded PhD Project (Students Worldwide)
About the Project
Fibrillin is an essential component of the extracellular matrix of all mammalian elastic tissues such as blood vessels, lung, joints and skin. Fibrillin gives our tissues elasticity and is also essential for cell signalling, storing TGFβ family growth factors within the matrix to provide a tissue store which is critical for normal development and tissue maintenance. When fibrillin is disrupted, as seen in Marfan syndrome and other diseases caused by fibrillin mutations, there is an increase in active TGFβ which results in widespread tissue damage. This paradox is not currently understood but indicates that in the absence of fibrillin, TGFβ can be more readily activated. TGFβ is activated by integrin cell surface receptors which also bind to fibrillin at the cell surface. However, it is not understood what role fibrillin plays in the process of TGFβ activation, therefore, we wish to understand the role of fibrillin in controlling the bioavailability and activation of TGFβ.
To address this question, complexes of fibrillin with latent TGFβ and integrins will be formed for structural and biochemical analysis. This project will use protein expression systems to purify specific protein complexes and then use electron microscopy techniques (CryoEM) to determine their 3-dimensional structures. These approaches will be supported by cell-based TGFβ signalling assays. State-of-the-art imaging approaches will be used, including cryo-electron microscopy where following the recent “Resolution Revolution”, atomic resolution structures from cryoEM data are now achievable, making previously intractable proteins such as extracellular matrix proteins accessible targets for structure determination. To aid in complex formation and structural analysis, this project will use single-domain antibody fragments (’nanobodies’) which are especially useful tools in protein structural biology, facilitating studies of conformationally dynamic proteins. An in vitro yeast surface display platform will be used to pan for nanobodies against fibrillin to aid the structural, biochemical and cell biology studies.
This project will determine whether integrin-TGFβ binding is enhanced or inhibited when latent TGFβ is bound to fibrillin using binding assays and cryoEM. Analysis of these structures will provide insight into the role of fibrillin in the storage and activation of TGFβ. Expression systems to purify latent TGFβ, fibrillin and the extracellular domain of integrin αVβ6 are already available in the lab for these studies. These interaction analyses will be complemented by signalling assays using a nano-Luciferase reporter assay for TGFβ signalling which is also available within the lab for this project.
https://www.wellcome-matrix.org/people/clair-baldock/
http://www.manchester.ac.uk/research/Clair.Baldock/
http://www.manchester.ac.uk/research/Alan.Roseman/
https://www.research.manchester.ac.uk/portal/thomas.a.jowitt.html
Entry Requirements:
Applicants must have obtained, or be about to obtain, at least an upper second class honours degree (or equivalent) in a relevant subject.
UK applicants interested in this project should make direct contact with the Principal Supervisor to arrange to discuss the project further as soon as possible. International applicants (including EU nationals) must ensure they meet the academic eligibility criteria (including English Language) as outlined before contacting potential supervisors to express an interest in their project. Eligibility can be checked via the University Country Specific information page (https://www.manchester.ac.uk/study/international/country-specific-information/).
If your country is not listed you must contact the Doctoral Academy Admissions Team providing a detailed CV (to include academic qualifications – stating degree classification(s) and dates awarded) and relevant transcripts.
Following the review of your qualifications and with support from potential supervisor(s), you will be informed whether you can submit a formal online application.
To be considered for this project you MUST submit a formal online application form - full details on how to apply can be found on the BBSRC DTP website www.manchester.ac.uk/bbsrcdtpstudentships
Funding Notes
Funding will cover UK tuition fees/stipend only. The University of Manchester aims to support the most outstanding applicants from outside the UK. We are able to offer a limited number of scholarships that will enable full studentships to be awarded to international applicants. These full studentships will only be awarded to exceptional quality candidates, due to the competitive nature of this scheme.
Equality, diversity and inclusion is fundamental to the success of The University of Manchester, and is at the heart of all of our activities. The full Equality, diversity and inclusion statement can be found on the website https://www.bmh.manchester.ac.uk/study/research/apply/equality-diversity-inclusion/
References
1: Lockhart, M, Newandee, H, Thomson, J, Weiss, AS, Baldock, C*, Tarakanova, A*.Transglutaminase-mediated cross-linking of tropoelastin to fibrillin stabilises the elastin precursor prior to elastic fibre assembly. J Mol Biol. 2020. In Press. doi: 10.1016/j.jmb.2020.08.023.
2: Thomson J, Singh M, Eckersley A, Cain SA, Sherratt MJ, Baldock C. Fibrillin microfibrils and elastic fibre proteins: Functional interactions and extracellular regulation of growth factors. Semin Cell Dev Biol (2019) 89:109-117.
3: Godwin, A., Starborg, T., Smith, D., Sherratt, M., Roseman, AM. & Baldock, C. (2018). Multiscale Imaging Reveals the Hierarchical Organisation of Fibrillin Microfibrils. J Mol Biol 430:4142-4155.
4: Troilo H, Steer R, Collins RF, Kielty CM, Baldock C. Independent multimerization of Latent TGFβ Binding Protein-1 stabilized by cross-linking and enhanced by heparan sulfate. Sci Rep (2016) 6:34347.
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