A new Tetrahymena actin-binding protein is localized in the division furrow.
Using an F-actin affinity column, a 60 kDa fragment of a 71 kDa F-actin-binding protein was partially purified from Tetrahymena pyriformis. After digestion of the 60 kDa fragment with cyanogen bromide, the N-terminal 21-amino acid sequence of one of the resulting peptides was found to show sequence similarity to a region near the actin-binding site (amino acid residues 260-281) of yeast fimbrin. An antibody prepared against a synthesized 21-mer oligopeptide reacted with the 71 kDa proteins in T. pyriformis and T. thermophila cell extracts, suggesting that the 60 kDa fragment was produced from the 71 kDa protein through partial digestion occurring during isolation. The 60 kDa fragment bound to Tetrahymena F-actin as well as to rabbit skeletal muscle F-actin, and induced the bundling of Tetrahymena F-actin. Indirect immunofluorescence revealed colocalization of the 71 kDa protein and actin in the oral apparatus and the deep fiber bundles in T. pyriformis. On the other hand, in T. thermophila, the 71 kDa protein was localized in the oral apparatus and the contractile vacuole pores during the interphase. During cytokinesis, the 71 kDa protein was localized in the division furrow. Therefore, the 71 kDa protein seems to associate with the actin cytoskeleton, and to regulate the actin filament organization during phagocytosis and cytokinesis in Tetrahymena.
Watanabe
A
A
Institute of Biological Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-0006.
Kurasawa
Y
Y
Watanabe
Y
Y
Numata
O
O
D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D002455:Cell Division; D019084:Fluorescent Antibody Technique, Indirect; D006868:Hydrolysis; D007399:Interphase; D008840:Microfilament Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D013769:Tetrahymena pyriformis / Q000166:cytology / Q000378:metabolism*
D000199:Actins; D008840:Microfilament Proteins; D010446:Peptide Fragments