INCREASED LEVELS OF MESSENGER-RNAS FOR TUBULIN AND OTHER FLAGELLAR PROTEINS AFTER AMPUTATION OR SHORTENING OF CHLAMYDOMONAS FLAGELLA
INCREASED LEVELS OF MESSENGER-RNAS FOR TUBULIN AND OTHER FLAGELLAR PROTEINS AFTER AMPUTATION OR SHORTENING OF CHLAMYDOMONAS FLAGELLA
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DOI：
10.1016/0092-8674(80)90633-9
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发表时间：
1980-01-01
期刊：
影响因子：
64.5
通讯作者：
ROSENBAUM, JL
中科院分区：
文献类型：
--
作者：
LEFEBVRE, PA;SILFLOW, CD;ROSENBAUM, JL
鞭毛蛋白合成的刺激发生在衣原体鞭毛去除或实验诱导的鞭毛再吸收到细胞中。这种刺激涉及微管蛋白和许多其他鞭毛蛋白的mRNA水平的增加。在去纤维化或鞭毛吸收后，从细胞中分离总RNA和poly（A）RNA，然后在网织红细胞裂解物系统中翻译。翻译产物的二维凝胶分析表明，RNA指导的α-半乳糖苷酶的体外合成和β-微管蛋白和许多其它鞭毛蛋白在爆燃或鞭毛再吸收后增加。α-体外合成的微管蛋白在二维凝胶上不与成熟鞭毛α-微管蛋白共迁移。微管蛋白体内标记实验表明，主要的α-爆燃后细胞中合成的微管蛋白与主要的α-微管蛋白共同迁移，体外制备的微管蛋白，而不是用主要的α-微管蛋白。存在于鞭毛中的微管蛋白。鞭毛α-微管蛋白可能作为前体合成，并在组装成鞭毛之前经历翻译后修饰。微管蛋白和其他鞭毛蛋白的合成可以被特异性地抑制和刺激。用IBMX [异丁基甲基黄嘌呤]处理细胞，诱导鞭毛吸收，导致微管蛋白和其他鞭毛蛋白的可翻译mRNA水平显著降低，而不影响非鞭毛蛋白的mRNA水平。
Stimulation of synthesis of flagellar proteins occurs in Chlamydomonas following flagellar removal or experimentally induced resorption of the flagella into the cell. This stimulation involves an increase in levels of mRNA for tubulin and many other flagellar proteins. Total RNA and poly(A) RNA were isolated from cells after deflagellation or flagellar resorption, and were then translated in the reticulocyte lysate system. Two-dimensional gel analysis of the translation products demonstrates that the RNA-directed in vitro synthesis of .alpha.- and .beta.-tubulins, and a number of other flagellar proteins, increases after deflagellation or flagellar resorption. The .alpha.-tubulin synthesized in vitro does not co-migrate on 2-dimensional gels with mature flagellar .alpha.-tubulin. In vivo labeling experiments show that the major .alpha.-tubulin synthesized in the cell after deflagellation co-migrates with the major .alpha.-tubulin made in vitro, not with the major .alpha.-tubulin present in the flagella. Flagellar .alpha.-tubulin is probably synthesized as a precursor and undergoes post-translational modification before assembly into the flagella. Synthesis of tubulin and other flagellar proteins can be specifically inhibited and stimulated. Treatment of cells with IBMX [isobutyl methylxanthine], which induces flagellar resorption, causes a marked decrease in levels of translatable mRNA for tubulin and other flagellar proteins, without affecting levels of mRNA for nonflagellar proteins.