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Abstract
The synthesis of post-translationally modified tubulins was examined during Artemia
development. Tubulin, either purified to homogeneity or in cell-free extracts, was
blotted to nitrocellulose and probed with a panel of antibodies. When purified tubulin
was examined, tyrosinated tubulin underwent a large decrease as development progressed
and this was accompanied by the appearance of detyrosinated tubulin in samples from
organisms developed 24 hr. The inclusion of carboxypeptidase inhibitors had a small
effect on the relative amounts of tyrosinated and detyrosinated tubulins in 24-hr
preparations. The amount of alpha- and beta-tubulin in cell-free extracts of Artemia
either remained relatively constant during development or increased slightly. The
same result was obtained for acetylated and tyrosinated tubulin. Detyrosinated tubulin
first appeared in 24-hr cell-free extracts and was only post-translationally modified
tubulin to increase, relative to the total amount of tubulin, as the brine shrimp
developed. As revealed by immunofluorescence staining, detyrosinated tubulin occurred
in many cell types of developing nauplii and was prominently displayed in mitotic
figures. Artemia, a complex metazoan animal, is thus able to grow for an extended
period of time in the absence of detyrosinated tubulin. This isoform is however, synthesized
in early larvae and may be required for the development of elongated cells including
those which encircle the gut. Detyrosination remains as the only developmentally related
change observed for brine shrimp tubulin.