Characteristics of basal body cartwheel reassembly.
Cartwheel complexes reassembled in a fraction derived by treating isolated oral apparatuses from Tetrahymena with 1.0 M KCl for 12 h. Approximately 40% of the KCl-soluble protein reassembled into cartwheel complexes. The reassembly reaction was protein-concentration dependent, and reassembled cartwheels were stable at 3 degrees C. Sucrose gradient centrifugation resolved 3 high molecular mass protein complexes from the KCl-soluble fraction. Each of the 3 complexes has a different mass, but each contains the same 5 polypeptides, 2 of which are probably tubulins. When these complexes were removed from the KCl-soluble fraction by high speed centrifugation, cartwheel reassembly did not occur. The 5 polypeptides in the high molecular mass complexes were among several other polypeptides resolved from reassembled cartwheels by 2-dimensional gel electrophoresis. The high molecular mass complexes are probably essential for cartwheel formation. The electrophoretic data also show that several polypeptides in the KCL-soluble fraction do not appear to be incorporated into cartwheels. These polypeptides are probably non-essential for cartwheel formation.
Gavin
R H
RH
Department of Biology, Brooklyn College of the City University of New York 11210.
Duffus
W A
WA
Contard
P C
PC
D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D007525:Isoelectric Focusing; D008854:Microscopy, Electron; D010455:Peptides / Q000378:metabolism; D011189:Potassium Chloride / Q000494:pharmacology; D011506:Proteins / Q000378:metabolism; D013194:Staining and Labeling; D013696:Temperature; D013768:Tetrahymena / Q000187:drug effects / Q000648:ultrastructure*
D002135:Calcium-Binding Proteins; D010455:Peptides; D011506:Proteins; D011189:Potassium Chloride