Striated flagellar roots occur associated with the basal apparatus of flagellated or ciliated eukaryotic cells and have been observed in association with centrioles and primary cilia. Striated flagellar roots are contractile organelles. We have recently demonstrated that striated flagellar roots are composed in large part of a calcium-binding 20,000 mol. wt. contractile phosphoprotein. This protein shares many features of the parvalbumin/troponin c/calmodulin superfamily of calcium-binding proteins. We present preliminary findings which suggest that antigenic homologs of the striated flagellar root protein occur associated with the centrioles and mitotic spindle poles of diverse organisms, including mammalian cells. Our ultrastructural studies indicate that the fibrous material and their condensations (pericentriolar satellites) of mammalian centrosomes is composed at least in part of antigenic homologs of striated flagellar roots. These findings have led to the proposal that striated flagellar root homologs respresent 'primitive' motile systems that are simple in composition and undergoe contraction directly in response to calcium binding. If these ideas are correct they have important implications for our understanding of eukaryotic cell structure and contractile function. We propose to characterize the antigenic homologs of the striated flagellar root protein in mammalian cells. We propose to generate a library of monoclonal antibodies and to use these in epitope mapping, immunofluorescent and immunoelectron microscopic studies of striated flagellar root homologs in mammalian cells. In addition we will characterize calcium-binding to the protein and its proteolytic fragments and study the characteristics of flagellar root protein phosphorylation.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM035258-02
Application #
3287705
Study Section
Molecular Cytology Study Section (CTY)
Project Start
1985-08-30
Project End
1988-07-31
Budget Start
1986-08-01
Budget End
1987-07-31
Support Year
2
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Case Western Reserve University
Department
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106
Sanders, M A; Salisbury, J L (1994) Centrin plays an essential role in microtubule severing during flagellar excision in Chlamydomonas reinhardtii. J Cell Biol 124:795-805
Baron, A T; Suman, V J; Nemeth, E et al. (1994) The pericentriolar lattice of PtK2 cells exhibits temperature and calcium-modulated behavior. J Cell Sci 107 ( Pt 11):2993-3003
Errabolu, R; Sanders, M A; Salisbury, J L (1994) Cloning of a cDNA encoding human centrin, an EF-hand protein of centrosomes and mitotic spindle poles. J Cell Sci 107 ( Pt 1):9-16
Harper, J D; Sanders, M A; Salisbury, J L (1993) Phosphorylation of nuclear and flagellar basal apparatus proteins during flagellar regeneration in Chlamydomonas reinhardtii. J Cell Biol 122:877-86
Goodenough, U W; Shames, B; Small, L et al. (1993) The role of calcium in the Chlamydomonas reinhardtii mating reaction. J Cell Biol 121:365-74
Baron, A T; Greenwood, T M; Bazinet, C W et al. (1992) Centrin is a component of the pericentriolar lattice. Biol Cell 76:383-8
Harper, J D; McCurdy, D W; Sanders, M A et al. (1992) Actin dynamics during the cell cycle in Chlamydomonas reinhardtii. Cell Motil Cytoskeleton 22:117-26
Coling, D E; Salisbury, J L (1992) Characterization of the calcium-binding contractile protein centrin from Tetraselmis striata (Pleurastrophyceae). J Protozool 39:385-91
Hulen, D; Baron, A; Salisbury, J et al. (1991) Production and specificity of monoclonal antibodies against calmodulin from Dictyostelium discoideum. Cell Motil Cytoskeleton 18:113-22
Baron, A T; Greenwood, T M; Salisbury, J L (1991) Localization of the centrin-related 165,000-Mr protein of PtK2 cells during the cell cycle. Cell Motil Cytoskeleton 18:1-14

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