MUSA - Alberta Pharmacy Students' Association
MUSA - Alberta Pharmacy Students' Association
MUSA - Alberta Pharmacy Students' Association
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University of <strong>Alberta</strong> Summer Students’ Research Day<br />
In 2011, over 200 undergraduate students participated in the Faculty of Medicine & Dentistry Summer Student<br />
Research Program. On October 15, 175 students presented posters at the 44th Annual Summer Students’ Research<br />
Day. Listed below are the 14 finalists from the poster competition. We congratulate the finalists and all participants.<br />
From these 14 finalists, two students were selected to represent the<br />
University of <strong>Alberta</strong> at the annual National Students’ Research<br />
Forum in Galveston, Texas. Their abstracts are presented below.<br />
Uncovering the role of topoisomerase II-beta<br />
binding protein 1 in dna replication stress response<br />
Mark Assmus, Charles Leung, Mark Glover<br />
DNA replication stress can lead to genomic instability which has<br />
been shown to be one of the primary hallmarks of cancer. TopBP1 is a<br />
crucial mediator protein found within the replication stress response<br />
in mammalian cells. TopBP1 activates Ataxia telangiectasia mutated<br />
related (ATR) kinase which phosphorylates many of the downstream<br />
substrates to initiate this response. The replication stress response<br />
involves specific interactions between the nine BRCA1 C terminus<br />
(BRCT) domains of TopBP1 and various proteins. More specifically,<br />
TopBP1 has been shown to provide an essential role in interacting<br />
with both ATR-interacting protein (ATRIP), Rad9-Rad1-Hus1 (9-1-1)<br />
complex as well as Mediator of DNA damage checkpoint protein 1<br />
(MDC1) which are all essential components of the response pathway.<br />
The crystal structure of TopBP1 BRCT 4/5 in complex with MDC1 was<br />
previously solved in our lab. The structure shows a unique mode of<br />
TopBP1 binding to MDC1 that involves the dimerization of two BRCT<br />
4/5 molecules. In an effort to further examine this interaction, I used<br />
a fluorescence polarization (FP) binding assay involving an MDC1<br />
FITC labelled di-phospho-peptide. I was able to express and purify<br />
GST fusion proteins of TopBP1 BRCT 4/5 and TopBP1 BRCT 5, as well<br />
as TopBP1 BRCT 5 alone, which were used for further FP studies.<br />
The results of the FP assays indicated that it is the BRCT 5 binding<br />
pocket which is primarily responsible for the interaction with MDC1<br />
and that the dimerization induced by GST allows for tighter binding.<br />
Additionally, mutant constructs of the putative BRCT 5 binding<br />
pocket were designed, successfully over-expressed and purified.<br />
The FP assays showed decreases in binding affinity associated with<br />
mutation of key conserved residues in the binding pocket. FP was<br />
also used to confirm that the phosphorylation of the MDC1 peptide<br />
is essential for TopBP1 BRCT 4/5 recognition. Taken together, these<br />
FP results further support the unique dimerization-based binding<br />
mechanism suggested by the crystal structure.<br />
PaX3 expression in melanoma<br />
Zachary Tan and D. Alan Underhill<br />
The transcription factor PAX3 is critical for development of neural<br />
crest lineages including melanocytes. Prior to birth, PAX3 is<br />
required for the proliferation of melanocyte precursors and it is<br />
thought to maintain an ‘undifferentiated plastic state’ in epidermal<br />
melanocytes after birth, as well in melanocyte stem cells. In<br />
addition, PAX3 is expressed throughout melanoma progression,<br />
from nevi to metastatic disease. Nevertheless, little is known about<br />
how PAX3 carries out these diverse roles. PAX3 is reported to be<br />
phosphorylated by Glycogen Synthase Kinase 3ß (GSK3ß). In the<br />
present study, the potential role of this kinase in modulating PAX3<br />
activity in B16F10 melanoma cells was examined using chemical<br />
inhibitors. Fluorescence Activated Cell Sorting (FACS) was used<br />
to assess cell cycle distribution and PAX3 levels were monitored<br />
by immunoblotting. Treatment of cells with the GSK3ß inhibitors<br />
lithium chloride (LiCl) or BIO caused decreased cell proliferation<br />
(P=0.05) and G2/M accumulation (P=0.05), and was associated<br />
with increased PAX3 expression (P=0.05). In contrast, knockdown<br />
of PAX3 using siRNA resulted in G1 accumulation (P=0.05).<br />
Immunofluorescence techniques for exogenous BrdU incorporation<br />
and endogenous PS10H3 allowed for direct microscopic visualization<br />
and quantification of cells in S and G2/M phase respectively. Upon<br />
PAX3 knockdown, there was significantly less BrdU incorporation<br />
and PS10H3 staining (P=0.05). Lastly, cell motility assays were<br />
conducted using live-cell Differential Interference Contrast (DIC)<br />
microscopy and analyzed using T-Scratch software. Interestingly,<br />
inhibition of GSK3ß as well as PAX3 knockdown was associated<br />
with markedly decreased cellular motility and proliferation. These<br />
investigations identify GSK3ß and as an important modulator of<br />
PAX3 levels in melanoma cells, and also suggest broader roles for<br />
PAX3 in regulating the G1 to S-phase transition in melanoma.<br />
Student Poster Title Supervisor<br />
Mark Assmus Uncovering the role of topoisomerase II-beta<br />
binding protein 1 in DNA replication stress<br />
response<br />
Christopher<br />
Beavington<br />
Department/<br />
Division<br />
Dr. Mark Glover Biochemistry<br />
University of <strong>Alberta</strong> Health Sciences Journal • April 2012 • Volume 7 • Issue 1 3<br />
Alanna<br />
Chomyn<br />
The structural studies of bacterial lactoferrin<br />
binding protein B from Neisseria meningitides<br />
Isolation of trkA expressing and IB4-binding<br />
sensory neurons through the use of saporin<br />
Nicholas Chua A model system for complex redox enzyme<br />
maturation<br />
Alexandru<br />
Cojocaru<br />
Michelina<br />
Kierzek<br />
Using inhibition of protein N-myristoylation<br />
towards the design of a synthetically lethal<br />
treatment of B-cell lymphomas<br />
Elucidating the molecular mechanisms<br />
of heart disease-linked mutations of<br />
phospholamban<br />
Stephanie Mah Capase 1 Inhibition in inflammatory bowel<br />
disease reduces epithelial cell extrusion<br />
Scott Meyer Investigating the quinone binding site of<br />
Escherichia coli fumarate reductase<br />
Robyn Millott The novel interaction between<br />
N-myristoyltransferase 1 and calnexin<br />
Kian Parseyan Proposed improvements for intraspinal<br />
microstimulation array fabrication and<br />
insertion<br />
Amit Persad Expression of ST8Sia family in developing<br />
chick retina and their role in AP2deltamediated<br />
axonal generation<br />
Raheem<br />
Suleman<br />
Does long life come from mom? Isolation of<br />
a longevity-conferring mitochondrial DNA<br />
mutation in Caenorhabditis elegans<br />
Dr. Joanne Lemieux Biochemistry<br />
Dr. Christine Webber Anatomy<br />
Dr. Joel H. Weiner Biochemistry<br />
Dr. Luc G.<br />
Berthiaume<br />
Cell Biology<br />
Dr. Howard S. Young Biochemistry<br />
Dr. Julia Liu Medicine/<br />
Gastroenterology<br />
Dr. Joel H. Weiner Biochemistry<br />
Dr. Marek Michalak Biochemistry<br />
Dr. Vivian K.<br />
Mushahwar<br />
Cell Biology<br />
Dr. Roseline Godbout Oncology<br />
Dr. Bernard D.<br />
Lemire<br />
Biochemistry<br />
Zachary Tan PAX3 expression in melanoma Dr. Alan Underhill Oncology<br />
Terri Waller Partial deficiency of adipose trigylceride<br />
lipase (ATGL) does not protect against<br />
diabetes-induced cardiac dysfunction<br />
Dr. Jason R.B. Dyck Pediatrics<br />
RESEARCH