In this project you will investigate how the altered nucleo-cytoskeletal architecture in aging cells and Hutchinson-Gilford Progeria Syndrome (HGPS) affects cell matrix adhesions and leads to nuclear import defects that cause HGPS symptoms. The nuclei of cells are directly linked to their extracellular environment through a bridge which encompasses focal adhesions, the actin cytoskeleton and the LINC complex. This bridge can act as a conduit via which extracellular forces are channelled to the nuclear interior and the Lamin A/C nucleo-cytoskeletal network, triggering changes in cellular behaviour and transcription. HGPS is a degenerative premature ageing disease resulting from de novo point mutations within the LMNA (Lamin A/C) gene that cause an alternate transcript called Progerin. Progerin expression affects nuclear morphology and is known to inhibit nuclear import/export of proteins, through clustering of nuclear pore complexes. Defects in nuclear export have recently been shown to be causative of the cellular phenotypes seen in HGPS. Ageing cell nuclei show similarities with HGPS nuclei for unknown reasons, possibly through a slow enrichment of Progerin over time.
1. Investigate effect of impaired nuclear force coupling on matrix adhesion dynamics and composition.
2. Interrogate effect of matrix adhesion complexes on nuclear pore complex function and nuclear protein import/export.
We have developed a novel BIoID2 based mass spectrometry workflow that enables us to investigate the molecular composition of cell matrix adhesion sites in 3D matrix environments. You will use established constructs for a proteomic profile of 3D matrix adhesions of Progerin or wildtype LaminA/C expressing cells.
You will perform an extensive analysis of adhesion dynamics in the cell lines available to us. You will use traction force microscopy to complement the Talin tension sensor experiments and evaluate force transduction of wildtype versus Progerin expressing cells. Together with the mass spectrometry this will yield a comprehensive analysis of the matrix adhesion changes in Progerin expressing ageing cells.
We have adapted atomic force microscopy-based techniques to evaluate the distribution and activity of nuclear pore complexes (NPCs) under controlled force application. You will investigate the NPC response of isolated age matched and progeria patient cell nuclei on force application. In parallel, you will investigate NPC distribution using superresolution microscopy techniques (STORM; SIM).
Together, this project will define the matrix adhesion site alterations caused by changes in the nucleo-cytoskeleton of HGPS patient and aged cell fibroblasts. It will show how reduced force transmission on the nucleus results in nuclear import/export defects and attempt to rescue the disease-causing nuclear export defects by altering matrix adhesion properties.
HOW TO APPLY
Applications should be made by emailing [email protected]
with a CV (including contact details of at least two academic (or other relevant) referees), and a covering letter – clearly stating your first choice project, and optionally 2nd and 3rd ranked projects, as well as including whatever additional information you feel is pertinent to your application; you may wish to indicate, for example, why you are particularly interested in the selected project(s) and at the selected University. Applications not meeting these criteria will be rejected.
In addition to the CV and covering letter, please email a completed copy of the Additional Details Form (Word document) to [email protected]
. A blank copy of this form can be found at: https://www.nld-dtp.org.uk/how-apply
Informal enquiries may be made to [email protected]
This is a 4 year BBSRC studentship under the Newcastle-Liverpool-Durham DTP. The successful applicant will receive research costs, tuition fees and stipend (£15,009 for 2019-20). The PhD will start in October 2020. Applicants should have, or be expecting to receive, a 2.1 Hons degree (or equivalent) in a relevant subject. EU candidates must have been resident in the UK for 3 years in order to receive full support. Please note, there are 2 stages to the application process.
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