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Universidade Nacional da Irlanda em Galway -- Ciência sem Fronteiras
PhD Project Template
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Use one form per project
Please complete & submit to international@nuigalway.ie as soon as possible, and by 27/11/2012
In your email, begin the subject line with [SWB] (be sure to use square brackets) to ensure that
your email is filed correctly. Emails will be automatically filed
PI name & contact details:
School:
Dr. Michael P. Carty
Michael.carty@nuigalway.ie
School of Natural Sciences, Biochemistry, NUI Galway
Has project been agreed with head (or
nominee) of proposed registration
school?
yes
Research Centre / group
affiliation:
Research group / centre
website:
Centre for Chromosome Biology
DNA Damage Response laboratory, Centre for Chromosome
Biology
http://www.chromosome.ie/researchers/carty/
PI website / link to CV
http://www.nuigalway.ie/biochemistry/staff/carty/index.html
Brief summary of PI research / research group / centre activity (2 or 3 lines max):
The main aim of the DNA Damage Response group led by Dr. Carty is to elucidate the molecular
basis of the response of human primary cells and cancer cells to DNA damaging agents, with a
specific interest in cancer therapeutics such as platinum-based drugs that interfere with DNA
replication.
Title & brief description of PhD project (suitable for publication on web):
A novel role for DNA damage-induced RPA modification in mitosis and cytokinesis.
Despite advances in the development of targeted therapies, DNA damaging agents including ionising
radiation and platinum-based drugs remain the mainstays of cancer treatment. Drugs such as
cisplatin act by inducing DNA lesions that block replication and transcription. Cells respond to
replication arrest by activation of a complex biochemical pathway termed the DNA damage
response (DDR). Targeting key components of the DDR could lead to the development of novel
therapeutics that potentiate the effects of DNA damaging agents used to treat cancer.
We have extensively characterised the DDR activated in response to replication arrest in model
human cell lines lacking DNA polymerase eta. These cells are deficient in bypass of replicationblocking lesions induced by UV radiation or cisplatin (1-5). Replication arrest activates PIK kinasemediated signalling, resulting in phosphorylation of a large number of downstream substrates,
modulating the outcome of exposure, including cell cycle arrest, DNA repair and apoptosis.
Replication protein A (RPA) the main single-stranded DNA binding protein in human cells is a key PIK
kinase substrate. RPA plays a critical role in preventing degradation of single-stranded DNA
generated during DNA replication or repair, and in recruiting repair complexes to damage sites. In
response to DNA damage, the RPA2 subunit of the trimeric RPA complex is phosphorylated on a
number of N-terminal sites, regulating the activity of the protein.
In the present project, a novel role for DNA damage-induced phosphorylation of RPA2 during
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Universidade Nacional da Irlanda em Galway -- Ciência sem Fronteiras
mitosis and cytokinesis will be investigated in detail, using cell biological and biochemical
approaches. Using immunofluorescence microscopy, it has been found that RPA2 phosphorylated
on serines 4 and 8, is relocalised to specific sites in human cells undergoing mitosis and cytokinesis.
To understand this process, the effect of mutating serines 4 and serine 8 to alanine on RPA2
localisation will be investigated using fluorescence microscopy and live cell imaging of GFP-tagged
RPA2 constructs The functional role of relocalisation will be analysed by downregulation of
endogenous RPA2 using siRNA, followed by re-expression of transfected wild-type or mutated
RPA2. Post-translational modification of RPA2 protein in mitotic cells will be characterised using
phosphospecific antibodies in western blotting; by isolation of His-tagged RPA2 and by mass
spectrometry. The functional effects will be analysed using in vitro assays for the single-stranded
DNA binding and protein-protein interaction activities of RPA2.
This project builds on considerable research into the response of human cells to DNA damage, and
will provide insight into a novel role for RPA phosphorylation in human cells undergoing mitosis and
cytokinesis. Detailed understanding of this post-translational modification of RPA2 could lead to the
development of agents that target specific functions of the RPA complex in replication or mitosis,
with potential for cancer treatment.
The project will provide excellent training in cell biology, molecular biology and biochemistry, in a
larger research environment that is highly focused on key aspects of chromosome biology.
See references:
1. Glynn et al. DNA Repair (2006)
2. Cruet-Hennequart, Glynn et al., DNA Repair (2008)
3. Cruet-Hennequart et al. Cell Cycle (2009)
4. O’Meara et al. Cancer Genomics Proteomics (2010)
5. Cruet-Hennequart et al. Subcellular Biochem. (2010)
Unique selling points of PhD project in NUI Galway:
State-of-the-art facilities are available in Biochemistry, the Centre for Chromosome Biology,
including cell culture facilities, dedicated long wavelength UVA-irradiation facilities, flow cytometry,
fluorescence microscopy with live cell imaging, and systems for gene and protein analysis. Students
will have access to the NUI Galway Mass Spectrometry facility, located in the adjacent School of
Chemistry, including Orbitrap and Ion-trap instruments.
High-level training in the use of all equipment and procedures, and in data analysis will be provided.
Students will be part of the structured PhD programme in Biochemistry in the School of Natural
Sciences, focused on Biochemistry. As part of this programme, in addition to the research thesis,
students take a series of focused modules aimed at developing core professional skills and
knowledge in areas relevant to future career development.
The DNA Damage Response group has a strong record of graduating PhD students, and in research
publication.
This is a highly interdisciplinary project carried out in the DNA Damage Response laboratory, part of
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Universidade Nacional da Irlanda em Galway -- Ciência sem Fronteiras
the Centre for Chromosome Biology, a network of 11 research groups with high-level expertise in all
aspects of chromosome biology. In addition there are ongoing links with international centres,
including the IGH, Montpellier, France.
Students will have opportunities to present their results in the weekly seminar series of the Centre
for Chromosome Biology where feedback from faculty and researchers directly working in the field
provides an invaluable learning experience. Biochemistry and the Centre for Chromosome Biology
host a strong seminar programme, where invited national and international research leaders
present their latest results.
Name & contact details for project queries, if different from PI named above:
Please indicate the graduates of which disciplines that should apply:
Biochemistry, Biotechnology, Molecular Biology, Cell Biology
Ciência sem Fronteiras / Science Without Borders Priority Area:
Please indicate the specific programme priority area under which the proposed PhD project fits- choose only
one (tick box):
Engineering and other technological areas
Pure and Natural Sciences (e.g. mathematics, physics, chemistry)/Physical Sciences (Mathematics,
Physics, Chemistry, Biology and Geosciences)
Health and Biomedical Sciences / Clinical, Pré-clinical and Health Sciences
Information and Communication Technologies (ICTs), Computing
Aerospace
Pharmaceuticals
Sustainable Agricultural Production
Oil, Gas and Coal
Renewable Energy
Minerals, Minerals Technology
Biotechnology
x
Nanotechnology and New Materials
Technologies for Prevention and Mitigation of Natural Disasters
Bioprospecting and Biodiversity
Marine Sciences
Creative Industry
New technologies in constructive engineering
Please indicate which of the following applies to this project (referring to Science Without Borders
arrangements):
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Suitable only as a Full PhD (Y/N): _ n (collaboration could also be considered)
Available to candidates seeking a Sandwich PhD arrangement (Y/N): __y___
Suitable for either/Don’t know: ___y__