Brunke K J, Young E E, Buchbinder B U, Weeks D P
Nucleic Acids Res. 1982 Feb 25;10(4):1295-310. doi: 10.1093/nar/10.4.1295.
During cell division and during the induction of tubulin synthesis that accompanies flagellar regeneration in Chlamydomonas reinhardi, four tubulin mRNAs of discrete molecular sizes are produced. During induction two beta tubulin mRNAs (2.47 kb and 2.34 kb) and two alpha tubulin mRNAs (2.26 kb and 2.13 kb) are synthesized in high abundance and in a closely coordinated fashion. Combined data from restriction enzyme mapping (i.e., Southern analysis) of genomic DNA and of Charon 30 recombinant clones bearing inserts of Chlamydomonas tubulin genes provide direct evidence for four distinct tubulin genes in this organism. Dot-blot analysis of the level of hybridization of a 32p nick-translated beta tubulin cDNA to genomic DNA from gametic cells and to a clone containing the beta 1 tubulin gene indicate that each beta 1 tubulin gene is present in one copy per cell. Additional hybridization experiments employing fragments of cDNA clones which selectively anneal to either the 3' or 5' portions of the two alpha tubulin genes or to one or both of the two beta tubulin genes suggest that each tubulin gene is actively transcribed to give rise to one of the four tubulin mRNAs. These observations further suggest that at most four basic types of tubulin subunits are produced by Chlamydomonas and that the heterogeneity of tubulin subunits reported to exist in the flagellar axoneme must arise as a result of post-translational modification.
在莱茵衣藻的细胞分裂以及鞭毛再生所伴随的微管蛋白合成诱导过程中,会产生四种分子大小不同的微管蛋白mRNA。在诱导过程中,两种β微管蛋白mRNA(2.47 kb和2.34 kb)以及两种α微管蛋白mRNA(2.26 kb和2.13 kb)以高丰度且紧密协调的方式合成。来自基因组DNA的限制性酶切图谱分析(即Southern分析)以及携带衣藻微管蛋白基因插入片段的Charon 30重组克隆的综合数据,为该生物体中四个不同的微管蛋白基因提供了直接证据。用32P缺口平移的β微管蛋白cDNA与配子细胞的基因组DNA以及含有β1微管蛋白基因的克隆进行杂交水平的斑点印迹分析表明,每个细胞中β1微管蛋白基因以单拷贝形式存在。使用cDNA克隆片段进行的其他杂交实验,这些片段可选择性地与两个α微管蛋白基因的3'或5'部分或两个β微管蛋白基因中的一个或两个退火,表明每个微管蛋白基因都被积极转录,从而产生四种微管蛋白mRNA之一。这些观察结果进一步表明,衣藻最多产生四种基本类型的微管蛋白亚基,并且据报道存在于鞭毛轴丝中的微管蛋白亚基的异质性必定是翻译后修饰的结果。