Abstract

The proposal under consideration focuses on the non-DNA-binding CBF(beta) subunit of the core binding protein CBF, a heterodimeric transcription factor. CBF(beta) is thought to allosterically regulate CBF(alpha), the DNA-binding subunit. The PI proposes to employ site directed mutagenesis to define the interaction surface in CBF(beta) to confirm the three dimensional structural information provided by NMR spectroscopy (Specific Aim 1). The NMR component of the research will be done in collaboration with Dr. John Bushweller at Dartmouth College. The investigator will also examine the expression of Cbfb during embryogenesis, determine the concentration of CBF-beta in hematopoietic cells (Specific Aim 2), and identify and biochemically characterize sequences in CBF-beta required for hematopoiesis (Specific Aim 3). This proposal is being submitted as an Interactive Research Project Grant in conjunction with J. Peter Gergen, who studies the Drosophila homologues of the CBF-beta protein. The two proposals describe a combined use of biochemical and genetic approaches to study the function of the CBF-beta protein in both the Drosophila and mouse systems.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA075611-03
Application #
2896191
Study Section
Hematology Subcommittee 2 (HEM)
Program Officer
Gallahan, Daniel L
Project Start
1997-07-15
Project End
2002-06-30
Budget Start
1999-07-01
Budget End
2000-06-30
Support Year
3
Fiscal Year
1999
Total Cost
Indirect Cost

  Institution

Name
Dartmouth College
Department
Biochemistry
Type
Schools of Medicine
DUNS #
041027822
City
Hanover
State
NH
Country
United States
Zip Code
03755

  Publications

Guo, Yalin; Maillard, Ivan; Chakraborti, Sankhamala et al. (2008) Core binding factors are necessary for natural killer cell development and cooperate with Notch signaling during T-cell specification. Blood 112:480-92
Talebian, Laleh; Li, Zhe; Guo, Yalin et al. (2007) T-lymphoid, megakaryocyte, and granulocyte development are sensitive to decreases in CBFbeta dosage. Blood 109:11-21
Li, Zhe; Lukasik, Steven M; Liu, Yizhou et al. (2006) A mutation in the S-switch region of the Runt domain alters the dynamics of an allosteric network responsible for CBFbeta regulation. J Mol Biol 364:1073-83
Li, Zhe; Yan, Jiangli; Matheny, Christina J et al. (2003) Energetic contribution of residues in the Runx1 Runt domain to DNA binding. J Biol Chem 278:33088-96
Zhang, Lina; Lukasik, Stephen M; Speck, Nancy A et al. (2003) Structural and functional characterization of Runx1, CBF beta, and CBF beta-SMMHC. Blood Cells Mol Dis 30:147-56
Speck, N A (2001) Core binding factor and its role in normal hematopoietic development. Curr Opin Hematol 8:192-6
Miller, J D; Stacy, T; Liu, P P et al. (2001) Core-binding factor beta (CBFbeta), but not CBFbeta-smooth muscle myosin heavy chain, rescues definitive hematopoiesis in CBFbeta-deficient embryonic stem cells. Blood 97:2248-56
Tang, Y Y; Shi, J; Zhang, L et al. (2000) Energetic and functional contribution of residues in the core binding factor beta (CBFbeta ) subunit to heterodimerization with CBFalpha. J Biol Chem 275:39579-88
Gu, T L; Goetz, T L; Graves, B J et al. (2000) Auto-inhibition and partner proteins, core-binding factor beta (CBFbeta) and Ets-1, modulate DNA binding by CBFalpha2 (AML1). Mol Cell Biol 20:91-103
Lewis, A F; Stacy, T; Green, W R et al. (1999) Core-binding factor influences the disease specificity of Moloney murine leukemia virus. J Virol 73:5535-47

Showing the most recent 10 out of 13 publications