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Matthew
Hildebrandt, B.Sc., Graduate Student |
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Research Project: DEAD
box 1 (DDX1) is a member of the DEAD box protein family of
putative RNA unwinding proteins. All DEAD box proteins share nine
conserved motifs. In addition, DDX1 has a SPRY domain implicated
in protein-protein interaction. DDX1 is co-amplified with MYCN
and overexpressed in a subset of retinoblastoma, neuroblastoma,
and rhabdomyosarcoma tumours. We propose that DDX1 plays a role
in the formation and/or progression of these tumours. |
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I am using a number of
approaches to examine DDX1 function. First, I am using the yeast
two-hybrid system to identify proteins that interact with DDX1.
Proteins that bind DDX1 will be further investigated by: (i)
determining how DDX1 interacting proteins bind to DDX1, (ii)
carrying out co-localization experiments to see whether DDX1 and
its interacting partners are found in the same subcellular
regions, and (iii) knocking-down levels of DDX1 interacting
proteins to study the effect on DDX1 levels, cell growth and
tumour formation. Second, I am generating a DDX1 inducible vector
to over-expressing DDX1 in mammalian cells. The goal of this
project is to determine whether growth properties and cell
morphology are affected. As well, I will investigate how DDX1
over-expression affects the levels, location and function of
proteins that bind DDX1. We are also using a knock-out mouse
model to elucidate the role of DDX1. Third, I will attempt to
generate the 3D structure of DDX1 using X-ray crystallography.
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Techniques: : Yeast 2-hybrid analysis, tissue
culture, siRNA transfection, western blot analysis, RT-PCR,
confocal microscopy, embryonic mouse manipulation (lacZ staining
and genotyping) |
Awards:
CIHR – CGSM; Department of Oncology Endowed Studentship; Walter H
Johns Graduate Fellowship
Abstracts/poster
presentations:
Hildebrandt, M., Katyal, S., Li, L., and Godbout, R.
Investigating and characterizing DEAD box protein 1 interacting
proteins. Alberta Cancer Research Institute Research Meeting, Banff,
Alberta (November 6-8, 2007).
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