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ECB-ART-35920
J Cell Sci 1994 Aug 01;107 ( Pt 8):2095-105. doi: 10.1242/jcs.107.8.2095.
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Centrosomal components immunologically related to tektins from ciliary and flagellar microtubules.

Steffen W , Fajer EA , Linck RW .


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Centrosomes are critical for the nucleation and organization of the microtubule cytoskeleton during both interphase and cell division. Using antibodies raised against sea urchin sperm flagellar microtubule proteins, we characterize here the presence and behavior of certain components associated with centrosomes of the surf clam Spisula solidissima and cultured mammalian cells. A Sarkosyl detergent-resistant fraction of axonemal microtubules was isolated from sea urchin sperm flagella and used to produce monoclonal antibodies, 16 of which were specific- or cross-specific for the major polypeptides associated with this microtubule fraction: tektins A, B and C, acetylated alpha-tubulin, and 77 and 83 kDa polypeptides. By 2-D isoelectric focussing/SDS polyacrylamide gel electrophoresis the tektins separate into several polypeptide spots. Identical spots were recognized by monoclonal and polyclonal antibodies against a given tektin, indicating that the different polypeptide spots are isoforms or modified versions of the same protein. Four independently derived monoclonal anti-tektins were found to stain centrosomes of S. solidissima oocytes and CHO and HeLa cells, by immunofluorescence microscopy. In particular, the centrosome staining of one monoclonal antibody specific for tektin B (tekB3) was cell-cycle-dependent for CHO cells, i.e. staining was observed only from early prometaphase until late anaphase. By immuno-electron microscopy tekB3 specifically labeled material surrounding the centrosome, whereas a polyclonal anti-tektin B recognized centrioles as well as the centrosomal material throughout the cell cycle. Finally, by immunoblot analysis tekB3 stained polypeptides of 48-50 kDa in isolated spindles and centrosomes from CHO cells.

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Genes referenced: dnah3 LOC100887844 LOC115919910 LOC594261 tubgcp2
???displayArticle.antibodies??? acetyl-LOC590586 Ab5 tekt1 Ab1 tekt1 Ab2 tekt1 Ab3 tekt1 Ab4 tekt1 Ab5 tekt2 Ab1 tekt2 Ab2 tekt2 Ab3 tekt2 Ab4 tekt2 Ab5 tekt2 Ab6 tekt4 Ab1 tekt4 Ab2