UNIVERSITY OF SUSSEX
GENOME DAMAGE & STABILITY CENTRE
The cellular responses to DNA damage:
A number of PhD positions are available within the Genome Damage and Stability
Centre. Students will join research-dominated environment with a well establish
international reputation. For more information: http://www.sussex.ac.uk/gdsc/
PhD stipends will be provided to undertake the research projects listed below. Potential
applicants should submit a letter of introduction and an accompanying CV to Mrs Gee
Wheatley at gdsc@sussex.ac.uk. The closing date for applications is the 28th Feb 2011.
Further information can be requested from the named supervisors.
UNIVERSITY OF SUSSEX
GENOME DAMAGE & STABILITY CENTRE
Role of the DNA damage response protein TDP2 in the cancer cell sensitivity to
clinical therapy
Prof Keith Caldecott, Dr Peter Schmid, Dr Tim Chevassut
To investigate the link between expression of the DNA repair proteins Tdp1 and Tdp2
and the sensitivity of cancer cells to chemotherapy. The project provides a unique
opportunity to combine aspects of both mechanistic and translational research to address
questions of direct relevance to clinical application. The project will be a collaboration
between the internationally renowned Genome Damage and Stability Centre and the
Brighton and Sussex Medical School and will provide expertise and training from both.
UNIVERSITY OF SUSSEX
GENOME DAMAGE & STABILITY CENTRE
The cellular responses to DNA damage:
Dr Hideo Tsubouchi
This project is focused on understanding the functional differences of mitotic and meiotic
homologous recombinases. The aims of the project are to isolate separation-of-function
alleles of RAD51 that show a defect specifically in mitotic recombination and investigate
what activity is defective in these separation-of-function rad51 mutants with the ultimate
aim of identifying the meiosis-specific function of Rad51 at the molecular level. This
project involves in vivo protein localisation analysis using fluorescent microscopy,
detection of recombination using genetic and molecular biological assays and
characterization of the properties of mutant homologous recombinases using biochemical
assays. This project provides an excellent opportunity to learn a wide spectrum of modern
biological techniques.
UNIVERSITY OF SUSSEX
GENOME DAMAGE & STABILITY CENTRE
The cellular responses to DNA damage:
Dr Jessica Downs
This project will investigate the Ies6 DNA binding subunit of the INO80 chromatin
remodeling complex in maintaining ploidy. Aneuploidy is a hallmark of tumours and
correlates with aggressiveness and malignancy. One commonly accepted mechanism of
aneuploidy is through endoreduplication, which leads to polyploidy followed by loss of
genetic information and increased chromosomal aberrations. We recently observed that
an Ino80 complex subunit, Ies6, is involved in regulating cell ploidy. We will explore the
mechanism by which Ies6 maintains normal ploidy using genetic suppressor screens of
ies6 mutant strains. In parallel, the effect of the DNA binding ability of Ies6 on the
activity and localization of the INO80 complex will be investigated. Finally, a
homologue of Ies6 is present in the mammalian INO80 complex, and the possibility that
human Ies6 maintains genome stability via a similar mechanism will be explored. This
project offers training in yeast molecular and cell biology coupled to experience with
mammalian cells.
UNIVERSITY OF SUSSEX
GENOME DAMAGE & STABILITY CENTRE
The cellular responses to DNA damage:
Prof Keith Caldecott, Dr Tony Oliver
This project is divided into two distinct but complementary sub-projects. For one part of
the project, the student will employ a variety of biochemical and cellular approaches to
identify novel protein partner/s of the XRCC1 BRCT1 domain, focusing initially on
specific candidates involved in DNA damage signalling/sensing (e.g. ATR/RAD9), DNA
synthesis (e.g. DNA polymerases such as epsilon & zeta), and structure specific
nucleases (e.g. the SLX4 nuclease scaffold protein). Subsequently, affinity-purification
and mass spectrometric analysis of epitope-tagged XRCC1 BRCT1 domain purified from
human cells will be used to identify novel partner proteins. For the second part of the
project, the student will address the structure of the complex formed between the XRCC1
BRCT2 domain and the BRCT domain of DNA ligase III. This BRCT-BRCT interaction
was amongst the first to be identified yet has still not yet been resolved at the structural
level. The project will provide a balance of biochemical, biophysical/structural, and
cellular approaches, providing a broad and competitive skill-set.
UNIVERSITY OF SUSSEX
GENOME DAMAGE & STABILITY CENTRE
The cellular responses to DNA damage:
Prof Tony Carr
This project will develop modifications to a site-specific replication fork arrest system
(Lambert et al, 2005) and optimise its analysis using chromatin immunoprecipitation
(ChIP) to understand replication fork restart (Lambert et al 2010), a process that
underpins genome stability and protects us from cancer. Once the systems are established,
the project will focus on creating specific epitope-tagged constructs in the fission yeast
genome (Watson et al, 2008) that represent the main replication complexes and proteins
(MCM, GINS, Polymerase holoenzymes etc) and use these in ChIP experiments to
monitor how and when these proteins association with replicating chromatin. Finally, a
series of experiments performed either with and without replication fork collapse and
restart at RTS1 will establish the proteins that are involved in replication following
replication restart. This project will provide a comprehensive training in molecular and
cellular biology and the use of model organisms in cancer research
UNIVERSITY OF SUSSEX
GENOME DAMAGE & STABILITY CENTRE
The cellular responses to DNA damage:
Dr Jo Murray
The aim of this project is to identify new hypomorphic and separation of function
mutants in the Smc5/6 complex components that will be used to shed light on the
function of Smc5/6 in replication fork stability. The project will use fission yeast and
Smc5/6 complex mutants defective in keeping stalled forks primed for restart will be
identified in a screen for mutants that suppress rDNA recombination. Mutations will
subsequently be correlated with the structure of the Smc5/6 complex to identify the
domains required. Selected mutants will be analysed by ChIP analysis for Rpa/Rad52
loading when replication is inhibited in HU. Sensitivity to transient HU or MMS
exposure, i.e. the consequences of a failure to load Rpa/Rad52 on replication resumption,
and the genetic dependencies will also be analysed. This analysis will correlate the
suppression of recombination, the recruitment of Rpa/Rad52 to stable stalled forks and
the domains/enzymatic activities of the Smc5/6 complex. In parallel representative
mutants will be analysed biochemically. Mutant Smc5/6 complexes will be purified using
myc-tagged Smc6. The domains identified will inform the choice of assays; e.g. ATP
hydrolysis, SUMO or ubiquitin ligase activity. The association of domains and the
stability of interaction will be investigated and confirmed by in vitro analysis. This
project will provide a broad training in molecular genetics, cell biology and biochemistry.