Abstract

The proposed research is aimed at bridging the gap of knowledge between the biochemistry, ultrastructure and cell physiology of cytoplasmic structure and contractility in non-muscle cells. The experiments take advantage of the developments from the last grant period in identifying actin-binding proteins, in developing quantitative fluoresence micropscopic assays, in proposing a working hypothesis on the relationship between cytoplasmic structure and contraction, and in implementing quantitative imaging techniques for basic questions in cell biology. The proposed experiment will continue the plan of integrating information obtained from motile cell extracts, reconstituted cell models, and from living cells. Changes in free [Ca++], pH, solation of the actin-based cytoskeleton, and contraction will be measured in in vitro models and in living cells. In addition, a critical test of the solation-contraction coupling hypothesis is proposed.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR032461-07
Application #
3156305
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1982-07-01
Project End
1991-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
7
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
Carnegie-Mellon University
Department
Type
Schools of Arts and Sciences
DUNS #
052184116
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213

  Publications

Vanni, Steven; Lagerholm, B Christoffer; Otey, Carol et al. (2003) Internet-based image analysis quantifies contractile behavior of individual fibroblasts inside model tissue. Biophys J 84:2715-27
Lagerholm, B Christoffer; Vanni, Steven; Taylor, D Lansing et al. (2003) Cytomechanics applications of optical sectioning microscopy. Methods Enzymol 361:175-97
Abraham, V C; Krishnamurthi, V; Taylor, D L et al. (1999) The actin-based nanomachine at the leading edge of migrating cells. Biophys J 77:1721-32
Burton, K; Park, J H; Taylor, D L (1999) Keratocytes generate traction forces in two phases. Mol Biol Cell 10:3745-69
DeBiasio, R L; LaRocca, G M; Post, P L et al. (1996) Myosin II transport, organization, and phosphorylation: evidence for cortical flow/solation-contraction coupling during cytokinesis and cell locomotion. Mol Biol Cell 7:1259-82
Giuliano, K A; Taylor, D L (1995) Measurement and manipulation of cytoskeletal dynamics in living cells. Curr Opin Cell Biol 7:4-12
Post, P L; DeBiasio, R L; Taylor, D L (1995) A fluorescent protein biosensor of myosin II regulatory light chain phosphorylation reports a gradient of phosphorylated myosin II in migrating cells. Mol Biol Cell 6:1755-68
Giuliano, K A; Post, P L; Hahn, K M et al. (1995) Fluorescent protein biosensors: measurement of molecular dynamics in living cells. Annu Rev Biophys Biomol Struct 24:405-34
Giuliano, K A; Taylor, D L (1994) Fluorescent actin analogs with a high affinity for profilin in vitro exhibit an enhanced gradient of assembly in living cells. J Cell Biol 124:971-83
Post, P L; Trybus, K M; Taylor, D L (1994) A genetically engineered, protein-based optical biosensor of myosin II regulatory light chain phosphorylation. J Biol Chem 269:12880-7

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