alpha4-Tubulin is involved in rapid formation of long microtubules to push apart the daughter centrosomes during earlyx Drosophila embryogenesis.
Although alpha4-tubulin comprises only about one-fifth of the alpha-tubulin pool in every Drosophila egg, in the absence of alpha4-tubulin - in eggs of the kavar(0)/- hemizygous females - only a tassel of short microtubules forms with two barely separated daughter centrosomes. We report that alpha4-tubulin is enriched in the long microtubules that embrace the nuclear envelope and suggest that they push apart daughter centrosomes along the nuclear perimeter during the initial cleavage divisions. In vitro tubulin polymerization showed that alpha4-tubulin is required for rapid tubulin polymerization. Since tubulin polymerization is slow inside eggs of the kavar(0)/- females, only short microtubules can form within the 4 to 5 minutes allowed for the process. A tassel of short microtubules with two barely separated centrosomes forms in every egg of the Kavar(18c)/+ females, in which the cytoplasm contains both wild-type and Kavar(18c)-encoded alpha4-tubulin with an E82K amino acid substitution (E82K-alpha4-tubulin). E82K-alpha4-tubulin is incorporated into the microtubules and renders them unstable. When injected into wild-type early cleavage embryos E82K-alpha4-tubulin slows down the formation of long microtubules and the separation of the daughter centrosomes. Surprisingly, when injected into late cleavage embryos E82K-alpha4-tubulin is non-toxic. Similarly, in the neuroblasts, ectopically expressed E82K-alpha4-tubulin becomes incorporated into the microtubules that grow sufficiently long and function normally.
Venkei
Zsolt
Z
Maternal Effect and Embryogenesis Research Group of the Hungarian Academy of Sciences at the University of Szeged, Faculty of Medicine, Department of Biology, Somogyi B. u. 4, H-6720 Szeged, Hungary.
Gáspár
Imre
I
Tóth
Gábor
G
Szabad
János
J
D000818:Animals; D002455:Cell Division / Q000502:physiology*; D002467:Cell Nucleus / Q000378:metabolism; D018385:Centrosome / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000166:cytology / Q000196:embryology; D005260:Female; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014404:Tubulin / Q000235:genetics / Q000378:metabolism*
D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; D014404:Tubulin