The function of nuclear envelope associated proteins in anchoring chromatin to the nuclear periphery

This is a jointly funded 3 years PhD studentship between Oxford Brookes University, UK, and Clermont University, France. The work will be divided evenly between the two labs (with periods of up to one year in both laboratories) and applicants must show an appropriate level of fluency in both French and English (IELTS 7.0 overall in English). The scientific focus of this project is to investigate the interactions between nuclear periphery components and heterochromatin in Arabidopsis thaliana.

Our mutual expertise in heterochromatin and the different components of the nuclear envelope provides this unique opportunity to study the relationship of chromatin organization and positioning at the nuclear envelope. During the past few years our groups and others identified severalnovel protein components localized to the nuclear periphery. These include SAD1/UNC84 (SUN) and KLARSICHT/ANC1/SYNE1 homology (KASH) proteins situated within the nuclear membrane (Graumann et al., 2010, Graumann et al., 2014), but also CROWDED NUCLEI (CRWN; Dittmer et al., 2007) and KAKU4 (Tamura et al., 2014) proteins localized underneath the nuclear membrane and believed to be orthologs of animal lamins. The Evans lab also recently identified a new class of nuclear envelope associated proteins, Nuclear Envelope Associated Proteins(NEAPs; Pawar-Menon, Graumann and Evans unpublished results). Using new methods to follow heterochromatin organization (Poulet et al., 2014) the Probst-Tatout lab showed that heterochromatin indeed localizes to the nuclear periphery (Poulet et al., in preparation) and characterized histone modifiers and chaperone complexes involved in heterochromatin organization (Ducet al., in press, Benoit et al., submitted).

Here, we propose to determine the interaction network between nuclear envelope associatedproteins using yeast two hybrid and in planta immuno-precipitation exploiting available specific antibodies and to identify the nuclear envelope components that establish a direct or indirect link with chromatin. Furthermore, we will use a combination of Chromatin Immunoprecipitation (ChIP) and deep sequencingto identify chromatin domains linked to the plant nuclear periphery. In addition, the proposed work aims to characterize expression and nuclear localization of a selection of these chromatin domains that interact with nuclear periphery components. As model tissues, we will use various Arabidopsis tissues including cotyledons during the first days post-germination, during which chromatin organization is highly dynamic and heterochromatic repetitive elements undergo clustering into chromocentres (Benoit et al., submitted). We will study these chromatin domains by RT-PCR, 3D-Fluorescence in situ hybridization (FISH), confocal microscopy and electron microscopy to determine the distribution of nuclear envelope components using available nuclear envelope markers. We further envisage analysing these aspects in chromatin modifier or remodeller mutants such as ddm1 or met1 that affect the spatial organization of nuclear domains (Feng et al., 2014 and Grob et al., 2014). Most of the proposed techniques such as the microscopy, RT-PCR, 3D-FISH, Y2H and Co-IP are established protocols in our labs.

Much of first year of this project will be carried out in GReD, Université Blaise Pascale, Clermont Ferrand, France under a co-tutelle agreement. The second and third years will be in the Sinclair Annex, Plant Cell Biology and Bioimaging group with periods of time at GReD.

As part of the studentship, any successful applicant will be required to undertake up to 6 hours undergraduate teaching per week during semesters without further remuneration. This may include supporting the teaching of statistics.
How to apply.

Eligibility: Only UK/EU nationals can apply (must be permanently resident in UK or EU) .


Please download application form-

http://www.hls.brookes.ac.uk/images/research/phd-studentship-application-form-jan-14.doc

Applications will only be accepted by email to the Research Administrator at the following address:[Email Address Removed]

Please additionally submit a CV and if appropriate IELTS test score certificate.

If you have any queries please contact Professor David Evans e-mail address [Email Address Removed]

PLEASE NOTE THAT YOU MUST NOT SUBMIT ANY APPLICATION-CV-IELTS TEST SCORE CERTIFICATE or QUERIES VIA FINDAPHD