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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:
Dr. Heinz Peter Nasheuer,
Biochemistry,
National University of Ireland, Galway
University Road
Galway
Ireland
Email: h.nasheuer@nuigalway.ie
phone: +353-91-49 2430
fax: +353-91-49 5504
School:
School of Natural Sciences
Has project been agreed with head (or
nominee) of proposed registration school?
Research Centre / group affiliation:
Research group / centre website:
PI website / link to CV:
Centre for Chromosome Biology;
Cell Cycle Control Group
http://www.chromosome.ie/researchers/nasheuer/
http://www.chromosome.ie/researchers/nasheuer/
Brief summary of PI research / research group / centre activity (2 or 3 lines max):
The research group has three major topics of interest:
 Cell cycle-dependent regulation and mechanisms of human DNA replication and repair pathways
 DNA replication of human polyoma viruses defining small molecules and novel small cellular
non-coding RNA as inhibitors of viral DNA replication
 Systems biology of protein-protein interactions in living cells using advanced microscopic
techniques
Title & brief description of PhD project (suitable for publication on web):
Mechanism of Cidofovir-derived antiviral agents inhibiting JC virus replication
The human JC virus or John Cunningham virus (JCV) belongs to the family of polyoma viruses which
are small, non-enveloped, icosahedral virus and its genome is organized in circular double-stranded
DNA. JCV causes Progressive Multifocal Leukoencephalopathy (PML), a demyelating disease of the
brain, in immunosuppressed humans. The viral replication is usually restricted to human glial cells.
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Universidade Nacional da Irlanda em Galway -- Ciência sem Fronteiras
Recently the JCV-caused disease got significant attention since PML was determined as side effect of
medical treatments for arthritis, Crohn’s Disease or multiple sclerosis (MS). During MS treatment
with the drug “Tysabri” the risk to develop PML is about 1/500 for patients treated but it rises to
1/85 in case of additional risk factors.
Recent findings suggest that the drug of Cidofovir inhibits viral replication but the mechanism is
unknown. Finding the mechanism for this drug may support the development of new drugs, which
will eliminate the PML side effect of treatments of MS patients, making these treatments safe for
more than 50% of humans who carry JCV. The proposed research will study the inhibition of viral
replication to prevent their reactivation in humans. Dr. Nasheuer’s group is a leading research
centre in the replication of human polyoma viruses.
In the project, DNA replication in cell-free systems and additional biochemical assays will be used to
determine the mechanism of Cidofovir in inhibiting JCV DNA replication in extracts derived from
human cells. They will be used to replicate viral origin-containing double-stranded DNA (dsDNA) in
the presence of JCV T antigen. Increasing amounts of Cidofovir (CDV) and its variants, CDV-P or CDVPP, will be added to the replication assays to determine their abilities of inhibit JCV DNA replication.
As controls for T antigen-independent DNA synthesis of replication extracts DNA synthesis on
primed M13 single stranded DNA will be carried out (DNA polymerase assay). These assays will be
performed in parallel with the same concentration of Cidofovir, CDV-P, and CDV-PP. Very high
concentration of CDV may inhibit DNA polymerases. The inhibition of DNA polymerases by
aphidicolin will act as a positive control for the inhibition of DNA polymerases. In case the viral DNA
replication is inhibited by Cidofovir, CDV-P or CDV-PP it will be important to determine the reaction
steps (origin-unwinding, initiation or elongation stage of DNA replication) that are sensitive to the
inhibitors. Here we will follow the stepwise procedure described by our lab.
In addition, the NTPase/helicase functions of T antigen will be analyses. The ATPase and DNA
helicase activity of JCV T antigen will be analysed using SV40 T antigen as a positive control. The
ATPase and DNA helicase activity are a requirement for the DNA replication function of JCV and
SV40 T antigen. Following these biochemical studies of T antigen we will determine whether the
enzyme activities of JCV T antigen are inhibited by Cidofovir and biochemically analyse the inhibiton
of Cidofovir and its variants.
Unique selling points of PhD project in NUI Galway:
NUI Galway projects should emphasise features that are not typically available in Brazil – specific equipment,
multi-disciplinarity, aspects of structured programme, links with industry, placements, links with other research
groups etc.
The student will gain experience in areas of genome stability including cell-free replication of viral
DNAs and analysis of inhibitors in biochemical reactions.
The PI is a member of the Centre for Chromosome Biology (CCB) at NUI Galway in Ireland and very
closely cooperates with research groups at NUI Galway and with international laboratories to
advance the knowledge in the mechanisms of genome instability.
http://www.chromosome.ie/
The Centre has gained international attention through its recent findings on the regulation of
trinucleotide repeats and the involvement of small noncoding RNA in the inhibition of viral DNA
replication.
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Universidade Nacional da Irlanda em Galway -- Ciência sem Fronteiras
CCB comprises a highly interactive network of researchers who are interested in topics focusing the
biology of genomes. The Centre currently consist of 11 groups with more than 90 researchers and
support staff. Their interests are focused on all molecular aspects underlying chromosome
replication, segregation, repair, evolution, chromatin regulation and transcription. To investigate
chromosome biology processes involved the Centre has available a range of technologies available
at the molecular, cellular and organismal level.
Name & contact details for project queries, if different from PI named above:
as above
Please indicate the graduates of which disciplines that should apply:
Graduates with a background molecular genetics/biology, biochemistry, cell biology, or related
disciplines should apply.
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
X
Sustainable Agricultural Production
Oil, Gas and Coal
Renewable Energy
Minerals, Minerals Technology
Biotechnology
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):
Suitable only as a Full PhD (Y/N):
Y