A predominant basic alpha-tubulin isoform present in prophase Xenopus oocyte decreases during meiotic maturation.
Xenopus oocytes are blocked in prophase of the first meiotic division. During the G2/M transition drastic changes occur both in the cytoskeletal organization and in the capacity of tubulin to polymerize. Posttranslational modification of tubulin isoforms might be one of the factors that control the dynamic properties of microtubules. We have therefore analysed, by two-dimensional polyacrylamide gel electrophoresis, the isotubulins purified from Xenopus oocytes, and we show that tubulin is resolved into at least four alpha-isoforms and four beta-isoforms. We have identified a basic alpha (alpha b)-tubulin isoform which is specific to prophase arrested oocyte and that progressively disappears during meiotic maturation; its decrease is initiated when the nuclear envelope breaks down and is controlled by the nucleus. Using 35S methionine labelled oocytes we demonstrate that the disappearance of the alpha b isotubulin results from both an arrest of its biosynthesis after maturation, and from posttranslational modification which induces a shift of this alpha-isoform to a more acidic pI. Moreover, in vitro experiments using 35S prelabelled tubulin purified from prophase oocytes show that metaphase extracts containing MPF activity are able to induce the acidification of the alpha b-isoform, suggesting that the observed posttranslational modification might be regulated by p34cdc2. However, the nature of this modification remains to be elucidated.
Thibier
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C
Laboratoire de Physiologie de la Reproduction, URA CNRS/INRA 1449, Université P et M Curie, Paris, France.
Denoulet
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P
Jessus
C
C
Ozon
R
R
D000818:Animals; D004527:Egg Proteins / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D007525:Isoelectric Focusing; D008540:Meiosis; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D011374:Progesterone / Q000494:pharmacology; D011418:Prophase; D014404:Tubulin / Q000145:classification / Q000378:metabolism*; D014982:Xenopus laevis / Q000378:metabolism
D004527:Egg Proteins; D014404:Tubulin; D011374:Progesterone