Two proteins crucial to proper B-cell development, the interferon regulatory factor (IRF) family member Pip and the ETS protein PU.1, must bind to each other and to specific DNA targets to exert their transcriptional control. PU.1 is a known oncogene and regulator of B-cell development. Members of the IRF family of proteins control processes in signal transduction, apoptosis, antiviral responses and oncogenesis. She wishes to study functional and structural aspects of their protein-protein interaction domains by an interdisciplinary approach using a combination of molecular biology and biophysical methods. First, she will ask what is the minimal protein interaction domain from Pip capable of forming ternary complexes with PU.1 and DNA. These studies will combine random and site-directed mutagenesis with an EMSA assay for complex formation and cotransfections for a functional assay. Second, she will determine what contribution each component makes to the stability of the ternary complex. These studies rely on surface plasmon resonance to determine kinetic and thermodynamic contributions from each macromolecule to the stability of ternary complexes. Finally she will use NMR to determine the solution structure of the minimal protein-protein interaction domain from Pip and a representative PEST domain from PU.1.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM056354-03
Application #
6386740
Study Section
Allergy and Immunology Study Section (ALY)
Program Officer
Chin, Jean
Project Start
1999-06-01
Project End
2003-05-31
Budget Start
2001-06-01
Budget End
2002-05-31
Support Year
3
Fiscal Year
2001
Total Cost
$315,167
Indirect Cost
Name
Sanford-Burnham Medical Research Institute
Department
Type
DUNS #
009214214
City
La Jolla
State
CA
Country
United States
Zip Code
92037
McKercher, Scott R; Lombardo, Christian R; Bobkov, Andrey et al. (2003) Identification of a PU.1-IRF4 protein interaction surface predicted by chemical exchange line broadening. Proc Natl Acad Sci U S A 100:511-6