Research Project: DEAD
box proteins represent a family of poorly understood RNA binding
and unwinding proteins that have been implicated in numerous
cellular processes. Our lab has identified a DEAD box protein,
called DEAD box 1 or DDX1, that is over-expressed in two
childhood tumours, retinoblastoma, a tumour of the eye, and
neuroblastoma, a tumour of the nervous system. Our goal is to
understand how expression of DDX1 affects the growth and
progression of both normal and tumour cells.
I currently have two projects
addressing the role of DDX1. The first project involves
identifying and characterizing the RNA molecules that naturally
bind to DDX1 in the cell. I am using two approaches for this
project. (1) RNA-DDX1 co-immunoprecipitations using two cell
lines: HeLa and retinoblastoma RB522A. Briefly, RNAs bound to
DDX1 are co-immunoprecipitated using an antibody to DDX1. The
RNAs are released, cDNA is synthesized and cloned into a vector.
The cDNAs are then sequenced and identified using DNA and protein
databases. (2) RNA immunoprecipitation – microchip arrays
(RIP-CHIP) which makes use of RNA microchip arrays to identify
RNAs bound to immunoprecipitated DDX1. The second project
involves using Drosophila as a model system for the study of DDX1
function. The advantage of using Drosophila is that many genetic
tools are available to modify DDX1 expression including
endogenous conditional knock-down of DDX1 using RNA interference,
tissue-specific knock-down of DDX1, generation of transgenic
flies over-expressing DDX1 is a tissue-specific manner.
microscopy, protein-RNA immunoprecipitation, RIP-CHIP, gel shift,
yeast-three-hybrid, Drosophila culture and dissection.
Germain, D., Li, L., Bleoo, S., Katyal, S. and Godbout, R. The
function and localization of DEAD box 1 (DDX1) in retinoblastoma cell
lines. Alberta Cancer Research Institute Research Meeting, Banff,
Alberta (November 6-8, 2007).
Germain, D., Hughes, S., Simmonds, A. and Godbout, R. Effects of Ddx1
on Drosophila melanogaster development. 4th Canadian Developmental
Biology Conference, Banff, Alberta (February 28 – March 2, 2008).
Germain, D., Hughes, S., Simmonds, A., Katyal, S. and Godbout, R. Investigating the role of DEAD box protein 1 (DDX1) in retinoblastoma. Alberta Cancer Research Institute Research Meeting, Banff, Alberta (November 25-27, 2008).
Germain, D., Hughes, S., Simmons, A. and Godbout, R. Characterization of the role of DDX1 in Drosophila melanogaster. 2nd Annual Canadian Human Genetics Conference, Harrison Hot Springs, BC (May 24-27, 2009).
Germain, D., Hughes, S., Simmons, A. and Godbout, R. Characterization of the role of DDX1 in Drosophila melanogaster. CANFLY 2009. The 10th Canadian Drosophila Research Conference, Jasper, Alberta (June 1-4, 2009).
Germain, D., Hughes, S., Simmonds, A and Godbout, R. Characterization of the role of DDX1 in Drosophila melanogaster development. 2009 Alberta Cancer Institute Research Meeting, Banff, Alberta (November 24-26, 2009).
Germain, D., Hughes, S., Simmonds, A. and Godbout, R. The Only Good Drosophila is a DEAD (Box1) Drosophila (actually The Only Good Fly is a DEAD Box 1. 5th Canadian Developmental Biology Conference, Mont-Tremblant, Quebec (April 8 – 11, 2010).
Germain, D.R., Graham, K., Glubrecht, D.D., Hughes, S., Simmonds, A., Mackey, J.R. and Godbout, R. DDX1, a novel independent prognostic marker of recurrence in breast cancer. 2010 Alberta Cancer Foundation Research Conference, Banff, Alberta (November 8-10, 2010).
Germain, D.R., Graham, K., Glubrecht, D.D., Hugh, J.C., Mackey, J.R. and Godbout, R. DEAD Box 1: A Novel and Independent Prognostic Marker for Early Recurrence in Breast Cancer. Breast Cancer Res. Treat. (in press).
Liu, R.-Z., Graham, K., Glubrecht, D.D., Germain, D.R., Mackey, J.R. and Godbout, R. Association of FABP5 expression with poor survival in triple-negative breast cancer: implication for retinoic acid therapy. Am. J. Pathol. (in press).
Alberta Cancer Foundation Studentship