The role of GEFs, potential regulators of LRRK2 GTPase activity, for the pathogenesis of Parkinson’s disease

Applications are invited for a 36 month PhD studentship starting in October 2010.

Project Description
Mutations in PARK8 encoding the cytosolic kinase LRRK2 are the most common known cause of familial and ‘idiopathic’ Parkinson’s disease. LRRK2 belongs to the ROCO family of proteins which are characterised by a Roc (Ras in complex proteins) domain with intrinsic GTPase activity and a COR (C-terminal of Roc) domain. The Roc-COR tandem domain controls LRRK2 kinase activity via an intramolecular process. The modification of LRRK2 GTPase and kinase activity by familial Parkinson’s disease mutations in the Roc, COR and kinase domains is believed to lead to neuronal cell death but the pathways involved remain elusive.

We have identified Guanine nucleotide exchange factors (GEFs) as interactors and potential regulators of LRRK2 GTPase activity. The PhD project includes characterization of the LRRK2 GEF interaction using: i) yeast two-hybrid and co-immunoprecipitation to depict protein interaction domains ii) dopaminergic differentiated cells to assess co-localisation and redistribution of LRRK2 and GEFs upon co-transfection iii) GTPase assays to assess the influence of GEF activity on LRRK2 GTPase activity iv) immunocytochemistry of primary neuronal cell cultures and immunohistochemistry of brain sections from LRRK2 knock-out, transgenic (harbouring Parkinson’s disease causing mutations) and normal control mice comparing GEF/LRRK2 distribution. This will allow conclusions on the physiological and pathological interplay of LRRK2 and GEFs. Techniques used will include: basis molecular biology, including cloning and mutagenesis; yeast two-hybrid; biochemistry e.g. co-immunoprecipitation; cell culture of immortalized cell lines and primary neuronal cell culture; transfection of mammalian cells; and GTPase assays.

This is a unique joint studentship between The School of Pharmacy and the University of Tübingen. The student will be primarily based at the School of Pharmacy in London but will spend approximately 50% of study at the University of Tübingen. This will allow the student to benefit from different facilities and expertise available to the supervisors. Upon successful completion of study, a PhD degree will be awarded from the School of Pharmacy, University of London.

Entry Requirements
Candidates must have, or expect to attain, a first or upper second class BSc degree in Biochemistry or Molecular Biology or a cognate discipline. Previous experience in cell biology, immunohistochemistry, microscopy or confocal imaging as well as spoken German would strengthen the application. Students whose first language is not English must pass the IELTS English proficiency test with a minimum score of 6.5 in each of the four skill areas.

Due to funding restrictions, only Home and EU students are eligible for the studentship. Overseas candidates are not eligible and regretfully enquiries and applications from ineligible candidates will not be acknowledged.

Stipend
The studentship covers tuition fees and a maintenance stipend for 36 months pegged to the BBSRC in London rate (approx £15,500).

Informal enquiries may be addressed to the supervisors:
Dr Kirsten Harvey
Department of Pharmacology
The School of Pharmacy
Tel +44 (0)20 7753 5888
Fax +44 (0)20 7753 5902
Email kirsten.harvey@pharmacy.ac.uk
Web profile http://www.pharmacy.ac.uk/kirsten_harvey.html

Dr Saskia Biskup
LRRK2 Group
University of Tübingen
Tel +49 70712972280
Fax +(49) 7071294522
Email saskia.biskup@medizin.uni-tuebingen.de

Applications
The application form is available to download at:
http://www.pharmacy.ac.uk/apply_phd.html

Candidates should send the completed application form, CV, personal statement explaining their interest in the project and the names and contact details (including email) of two academic referees to Lauren O’Brien, Registry Officer, The School of Pharmacy, University of London, 29/39 Brunswick Square, London WC1N 1AX, tel: 020 7753 5958, fax: 020 7753 5829, phd@pharmacy.ac.uk

The application closing date is 31 March 2010. The studentship will be held open after this date until a suitable candidate is appointed.