Injection of Antibodies to the Cytoskeleton Into Cells

Feramisco, James R.

Abstract

The structural and functional significance of the actin microfilament system in nonmuscle cells in cell motility and growth will be analyzed. Using biochemistry, cell biology, immunology, and molecular biology, an integrated approach to this problem will be followed. Through the production of highly specific monoclonal antibodies directed against three Ca2+ regulated microfilament components (alpha-actinin, vinculin and myosin light chain kinase) and the subsequent microinjection of these antibodies into living nonmuscle cells, the functional roles of these proteins will be determined. Through the microinjection of Ca2+/EGTA buffers into living cells, the role of Ca2+ in the dynamics of the microfilament system will be studied. And, through the molecular cloning of the genes encoding the biochemically distinct muscle and nonmuscle forms of both alpha-actinin and vinculin, the molecular basis for the different biochemical activities will be examined.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of General Medical Sciences (NIGMS)
• Type: Research Project (R01)
• Project #: 5R01GM028277-06
• Application #: 3275572
• Study Section: Cellular Biology and Physiology Subcommittee 1 (CBY)
• Project Start: 1980-07-01
• Project End: 1986-07-31
• Budget Start: 1985-08-01
• Budget End: 1986-07-31
• Support Year: 6
• Fiscal Year: 1985

Institution

• Name: Cold Spring Harbor Laboratory
• DUNS #: 065968786
• City: Cold Spring Harbor
• State: NY
• Country: United States
• Zip Code: 11724

Publications

Bar-Sagi, D; Feramisco, J R (1986) Induction of membrane ruffling and fluid-phase pinocytosis in quiescent fibroblasts by ras proteins. Science 233:1061-8

Welch, W J; Feramisco, J R (1985) Disruption of the three cytoskeletal networks in mammalian cells does not affect transcription, translation, or protein translocation changes induced by heat shock. Mol Cell Biol 5:1571-81

Catelli, M G; Binart, N; Jung-Testas, I et al. (1985) The common 90-kd protein component of non-transformed '8S' steroid receptors is a heat-shock protein. EMBO J 4:3131-5

Bar-Sagi, D; Feramisco, J R (1985) Microinjection of the ras oncogene protein into PC12 cells induces morphological differentiation. Cell 42:841-8

Feramisco, J R; Clark, R; Wong, G et al. (1985) Transient reversion of ras oncogene-induced cell transformation by antibodies specific for amino acid 12 of ras protein. Nature 314:639-42

Welch, W J; Feramisco, J R (1985) Rapid purification of mammalian 70,000-dalton stress proteins: affinity of the proteins for nucleotides. Mol Cell Biol 5:1229-37

Welch, W J (1985) Phorbol ester, calcium ionophore, or serum added to quiescent rat embryo fibroblast cells all result in the elevated phosphorylation of two 28,000-dalton mammalian stress proteins. J Biol Chem 260:3058-62

Welch, W J; Feramisco, J R; Blose, S H (1985) The mammalian stress response and the cytoskeleton: alterations in intermediate filaments. Ann N Y Acad Sci 455:57-67