Abstract

While considerable attention has been paid to the biochemical regulation of apoptosis, less effort has been directed towards elucidating the mechanisms that account for the morphology of apoptosis. This proposal will utilize fluorescent analogs of key cytoskeletal proteins to examine apoptosis-associated changes in cellular architecture in real time in living cells. Regulation by Rho family GTPases and cleavage of cytoplasmic components by caspases will be investigated. Particular attention will be paid to the role of D4GDI cleavage by CPP32 and how this alters Rho GTPase activity. Novel fluorescent indicators of protein cleavage will be developed to examine the kinetics and location of D4GDI and actin during apoptosis. Cytoskeletal control of specific Rho GTPases and their interactions in regulatory cascades will be determined by functional assays employing dominant negative and constitutively active mutants.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
5R01AG015430-03
Application #
6043106
Study Section
Human Embryology and Development Subcommittee 1 (HED)
Program Officer
Sierra, Felipe
Project Start
1997-09-01
Project End
2001-07-31
Budget Start
1999-08-01
Budget End
2000-07-31
Support Year
3
Fiscal Year
1999
Total Cost
Indirect Cost

  Institution

Name
Scripps Research Institute
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037

  Publications

Subauste, M Cecilia; Nalbant, Perihan; Adamson, Eileen D et al. (2005) Vinculin controls PTEN protein level by maintaining the interaction of the adherens junction protein beta-catenin with the scaffolding protein MAGI-2. J Biol Chem 280:5676-81
Subauste, M Cecilia; Pertz, Olivier; Adamson, Eileen D et al. (2004) Vinculin modulation of paxillin-FAK interactions regulates ERK to control survival and motility. J Cell Biol 165:371-81
Han, Wen-Ge; Liu, Tiqing; Himo, Fahmi et al. (2003) A theoretical study of the UV/visible absorption and emission solvatochromic properties of solvent-sensitive dyes. Chemphyschem 4:1084-94
Toutchkine, Alexei; Kraynov, Vadim; Hahn, Klaus (2003) Solvent-sensitive dyes to report protein conformational changes in living cells. J Am Chem Soc 125:4132-45
Hahn, Klaus; Toutchkine, Alexei (2002) Live-cell fluorescent biosensors for activated signaling proteins. Curr Opin Cell Biol 14:167-72
Kiosses, W B; Hahn, K M; Giannelli, G et al. (2001) Characterization of morphological and cytoskeletal changes in MCF10A breast epithelial cells plated on laminin-5: comparison with breast cancer cell line MCF7. Cell Commun Adhes 8:29-44
Schurmann, A; Mooney, A F; Sanders, L C et al. (2000) p21-activated kinase 1 phosphorylates the death agonist bad and protects cells from apoptosis. Mol Cell Biol 20:453-61
Subauste, M C; Von Herrath, M; Benard, V et al. (2000) Rho family proteins modulate rapid apoptosis induced by cytotoxic T lymphocytes and Fas. J Biol Chem 275:9725-33
Kraynov, V S; Chamberlain, C; Bokoch, G M et al. (2000) Localized Rac activation dynamics visualized in living cells. Science 290:333-7
Zenke, F T; King, C C; Bohl, B P et al. (1999) Identification of a central phosphorylation site in p21-activated kinase regulating autoinhibition and kinase activity. J Biol Chem 274:32565-73

Showing the most recent 10 out of 11 publications