This notification is posted on behalf of Prof Wim Hol. Interested parties
should reply to him at:
[EMAIL PROTECTED]
or at the address below.
Postdoctoral Fellow Structural Biology of the RNA editing editosome
JOB DESCRIPTION:
The project aims at understanding the structure and functioning of the
unique RNA-editing editosome from Trypanosoma brucei and related
trypanosomatid protozoan species. These organisms are the causative agents of a
variety of diseases in tropical and subtropical areas, including sleeping
sickness in Sub-Saharan Africa, Chagas disease in Latin America and
leishmaniasis throughout the tropics and subtropics. Several essential genes in
the mitochondria of these protozoa undergo a fascinating U-insertion/deletion
RNA editing process involving several protein and multi-protein complexes. The
RNA-editing editosome of about 1.5 million Daltons performs the actual editing
steps, involving an enzyme-cascade principle. The editosome consists of over 15
different proteins with most, if not all, of these proteins present in multiple
copies.
The goal of the project is to unravel, by molecular biology and
crystallographic approaches: (i) protein-protein and protein-RNA interactions
within the editosome; and, (ii) crystal structures of components and
sub-complexes of this multi-protein assembly in complex with a variety of RNA
molecules. Since several of the editosome proteins have been shown to be
essential for the parasites, the results obtained provide a platform for
structure-based drug design.
The successful candidate will have the opportunity to carry out (i)
molecular biology, protein expression and purification methods to obtain
insight into protein-protein and protein-RNA interactions involving the
editosome, and (ii) to determine high resolution crystal structures of
individual editosome components, and in particular of large sub-complexes of
the editosome with RNA. Numerous expression systems are already available for
preparing complexes containing multiple editosome proteins.
For further information regarding the editosome see:
Deng, J., Schnaufer, A., Salavati, R., Stuart, K. & Hol, W. G. J. (2004). High
Resolution Crystal Structure of an Editosome Enzyme from Trypanosoma brucei: RNA
Editing ligase I. J Mol Biol 343, 601-613
Deng, J., Lewis Ernst, N., Turley, S., Stuart, K. & Hol, W. G. J. (2005).
Structural basis for UTP specificity of RNA editing TUTases from Trypanosoma
brucei. EMBO J. 24, 4007-4017.
For information regarding the research in our lab see the websites:
http://www.bmsc.washington.edu/WimHol/
and
http://depts.washington.edu/biowww/faculty/hol.html
JOB REQUIREMENTS:
- Good knowledge and extensive experience with molecular biology for protein overexpression
- Experience with:
- protein expression and purification methods of soluble proteins in E
coli
- characterizing purified soluble proteins
- crystallization methods for proteins
- protein crystal cryo-protection procedures
- protein structure determination methods, including: X-ray
diffraction data collection and data processing; selenomethionine SAD and MAD
phasing and/or multiple isomorphous replacement and/or molecular replacement;
density modification; model building; crystallographic refinement; structure
analysis and structure validation procedures using multiple computer programs
and interactive graphics techniques.
- Excellent interpersonal skills to function optimally in the editosome project
team and to cooperate with collaborators in other institutions.
The Following experience would be a plus:
- Molecular biology for obtaining truncated protein variants and
implementing surface mutations to enhance the probability of crystal growth
- production of RNA
- protein expression and purification methods of soluble proteins in insect cells
START DATE: Immediately
INSTITUTION: Department of Biochemistry
Biomolecular Structure Center
School of Medicine
Box 357742
University of Washington
Seattle, WA, 98195 USA
