Much information has accumulated over the last 15 years regarding the molecular structure, assembly and cell biology of intermediate filaments (IFs). Over the past few years, cell biology, protein chemistry and molecular genetics have revealed an impressive body of information regarding the structure and assembly of IFs. The IF protein domains elucidated in the 1980's now appear to have important roles in forming and stabilizing filament structure. We now need to know what those roles are and how they are regulated in the cell. Dr. Ip proposes to continue his studies on the mechanisms of formation of IF protein tetrameres, higher order oligomers, filaments and filament networks, and to investigate how these abundant cytoskeletal elements interact functionally with other cellular components.
Four specific aims are proposed: (1) to analyze the tail domain of vimentin and its significance in IF assembly; (2) to determine the subunit composition of the neurofilament (NF) building block by in vitro studies, using bacterially expressed recombinant NF proteins; (3) to define specific interacting domains in vimentin IFs, using a two-hybrid system in yeast; and (4) to understand functional interactions between IFs and desmosomes, which together confer structure and stability to tissues. This information is of potential clinical relevance, given that point mutations in IF genes are known to be the basis of several skin diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR035973-11
Application #
2442797
Study Section
Special Emphasis Panel (ZRG2-BIOL-2 (01))
Project Start
1985-01-01
Project End
1999-06-30
Budget Start
1997-07-01
Budget End
1998-06-30
Support Year
11
Fiscal Year
1997
Total Cost
Indirect Cost
Name
University of Cincinnati
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
City
Cincinnati
State
OH
Country
United States
Zip Code
45221
Meng, J J; Ip, W (2001) A yeast two-hybrid approach for probing cytoskeletal protein interactions. Methods Mol Biol 161:255-68
Meng, J J; Bornslaeger, E A; Green, K J et al. (1997) Two-hybrid analysis reveals fundamental differences in direct interactions between desmoplakin and cell type-specific intermediate filaments. J Biol Chem 272:21495-503
Carpenter, D A; Ip, W (1996) Neurofilament triplet protein interactions: evidence for the preferred formation of NF-L-containing dimers and a putative function for the end domains. J Cell Sci 109 ( Pt 10):2493-8
Meng, J J; Khan, S; Ip, W (1996) Intermediate filament protein domain interactions as revealed by two-hybrid screens. J Biol Chem 271:1599-604
Meng, J J; Khan, S; Ip, W (1994) Charge interactions in the rod domain drive formation of tetramers during intermediate filament assembly. J Biol Chem 269:18679-85
Makarova, I; Carpenter, D; Khan, S et al. (1994) A conserved region in the tail domain of vimentin is involved in its assembly into intermediate filaments. Cell Motil Cytoskeleton 28:265-77
Mulligan, L; Balin, B J; Lee, V M et al. (1991) Antibody labeling of bovine neurofilaments: implications on the structure of neurofilament sidearms. J Struct Biol 106:145-60
Tao, J X; Ip, W (1991) Site-specific antibodies block kinase A phosphorylation of desmin in vitro and inhibit incorporation of myoblasts into myotubes. Cell Motil Cytoskeleton 19:109-20
Birkenberger, L; Ip, W (1990) Properties of the desmin tail domain: studies using synthetic peptides and antipeptide antibodies. J Cell Biol 111:2063-75
Ip, W; Saeed, T (1990) Desmin-binding specificities of two desmin CNBr fragments that correspond to the headpiece domain and Helix 1B. Cell Biol Int Rep 14:1119-27

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