The goal of this research project is to determine how proteins assemble into complexes that function to direct development and adaptive responses. The central focus is on LIM domains, Zn2+ binding protein modules located in both nuclear and cytoplasmic proteins, that act to assemble molecular complexes. LIM domain containing proteins are essential for nervous system, hematopoietic and muscle development; they are also essential for cytoskeletal function.
Specific aims i nclude: 1) Determine the structural basis of molecular recognition by LIM domains. Nuclear LIM interactor (NLI) is the major high affinity target recognized by LIM homeodomain (LH) and LIM only (LMO) proteins. Goals are to a) determine features of NLI involved in LIM domain binding, b) determine features of nuclear LIM domains necessary for high affinity molecular recognition, c) determine the kinetics and stoichiometry of NLI binding and d) determine the structure of a nuclear LIM domain NLI complex. 2. Analyze the function of nuclear LIM interactor. Predictions based on biochemical studies will be critically tested in functional assays. Goals are to a) analyze the function of NLI in retinoic acid-induced neuronal differentiation of P19 cells and b) analyze the function of NLI in transcription mediated by LH and LMO proteins. 3. Determine the function of nuclear LIM only proteins. These studies will focus on a newly discovered protein LM04. Goals are to a) determine the expression of LM04 during mouse development and compare it to LH, LMO and transcription factors essential for specific developmental processes. b) determine the effect of targeted disruption of LM04 in mouse development. 4. Analyze the function of cytoplasmic LIM domains. Many cytoplasmic LIM domain proteins regulate the cytoskeleton and have molecular recognition that is distinct from nuclear LIM domains. Goals are to a) identify and characterize high affinity interactions that involve cooperation between cytoplasmic LIM domains and b) determine the function of the LIM domains of LIM kinase. Regulation of LIM kinase that controls features of the actin cytoskeleton will be defined.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
2R01DK013149-32
Application #
2758332
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Program Officer
Sato, Sheryl M
Project Start
1977-01-01
Project End
2003-12-31
Budget Start
1999-01-01
Budget End
1999-12-31
Support Year
32
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of California San Diego
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Yeo, Michele; Lin, Patrick S (2007) Functional characterization of small CTD phosphatases. Methods Mol Biol 365:335-46
Lee, Soo-Kyung; Jurata, Linda W; Nowak, Roberta et al. (2005) The LIM domain-only protein LMO4 is required for neural tube closure. Mol Cell Neurosci 28:205-14
Yeo, Michele; Lee, Soo-Kyung; Lee, Bora et al. (2005) Small CTD phosphatases function in silencing neuronal gene expression. Science 307:596-600
Gill, Gordon N (2003) Decoding the LIM development code. Trans Am Clin Climatol Assoc 114:179-89
Yeo, Michele; Lin, Patrick S; Dahmus, Michael E et al. (2003) A novel RNA polymerase II C-terminal domain phosphatase that preferentially dephosphorylates serine 5. J Biol Chem 278:26078-85
Thaler, Joshua P; Lee, Soo-Kyung; Jurata, Linda W et al. (2002) LIM factor Lhx3 contributes to the specification of motor neuron and interneuron identity through cell-type-specific protein-protein interactions. Cell 110:237-49
van Meyel, D J; O'Keefe, D D; Thor, S et al. (2000) Chip is an essential cofactor for apterous in the regulation of axon guidance in Drosophila. Development 127:1823-31
Edwards, D C; Sanders, L C; Bokoch, G M et al. (1999) Activation of LIM-kinase by Pak1 couples Rac/Cdc42 GTPase signalling to actin cytoskeletal dynamics. Nat Cell Biol 1:253-9
van Meyel, D J; O'Keefe, D D; Jurata, L W et al. (1999) Chip and apterous physically interact to form a functional complex during Drosophila development. Mol Cell 4:259-65
Klingbeil, C K; Gill, G N (1999) A basic residue, Lys 782, composes part of the ATP-binding site on the epidermal growth factor receptor tyrosine kinase. Arch Biochem Biophys 363:27-32

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