Liaud M F, Brandt U, Cerff R
Institut für Genetik, Technische Universität Braunschweig, Germany.
Plant Mol Biol. 1995 May;28(2):313-25. doi: 10.1007/BF00020250.
We characterized a nuclear gene and its corresponding cDNA encoding beta-tubulin (gene TubB1) of the marine red alga Chondrus crispus. The deduced TubB1 protein is the most divergent beta-tubulin so far reported with only 64 to 69% amino acid identity relative to other beta-tubulins from higher and lower eukaryotes. Our analysis reveals that TubB1 has an accelerated evolutionary rate probably due to a release of functional constraints in connexion with a specialization of microtubular structures in rhodophytes. It further indicates that isoform diversity and functional differentiation of tubulins in eukaryotic cells may be controlled by independent selective constraints. TubB1 has a short spliceosomal intron at its 5' end which seems to be a characteristic feature of nuclear protein-coding genes from rhodophytes. The splice junctions of the four known rhodophyte introns comply well with the corresponding consensus sequences of higher plants in agreement with previous suggestions from phylogenetic inference that red algae and green plants may be sister groups. The paucity and asymmetrical location of introns in rhodophyte genes can be explained by differential intron loss due to conversion of genes by homologous recombination with cDNAs corresponding to reverse transcribed mRNAs or partially spliced pre-mRNAs, respectively. The identification of an intron containing TubB1 cDNA in C. crispus confirms that pre-mRNAs can escape both splicing and degradation in the nucleus prior to transport into the cytoplasm. Differential Southern hybridizations under non-stringent conditions with homologous and heterologous probes suggest that C. crispus contains a second degenerate beta-tubulin gene (or pseudogene?) which, however, is only distantly related to TubB1 as it is to the more conserved homologues of other organisms.
我们对一种海洋红藻皱波角叉菜的编码β-微管蛋白的核基因及其相应的cDNA(基因TubB1)进行了表征。推导的TubB1蛋白是迄今为止报道的分歧最大的β-微管蛋白,相对于来自高等和低等真核生物的其他β-微管蛋白,其氨基酸同一性仅为64%至69%。我们的分析表明,TubB1具有加速的进化速率,这可能是由于与红藻中微管结构特化相关的功能限制的释放。这进一步表明,真核细胞中微管蛋白的异构体多样性和功能分化可能受独立的选择限制控制。TubB1在其5'端有一个短的剪接体内含子,这似乎是红藻核蛋白编码基因的一个特征。四个已知的红藻内含子的剪接接头与高等植物的相应共有序列非常吻合,这与系统发育推断中红藻和绿色植物可能是姐妹群的先前建议一致。红藻基因内含子的稀少和不对称定位可以分别通过与对应于逆转录mRNA或部分剪接的前体mRNA的cDNA同源重组导致的基因转换引起的内含子差异丢失来解释。在皱波角叉菜中鉴定出一个含有TubB1 cDNA的内含子,证实了前体mRNA在转运到细胞质之前可以在细胞核中逃避剪接和降解。在非严格条件下用同源和异源探针进行的差异Southern杂交表明,皱波角叉菜含有第二个退化的β-微管蛋白基因(或假基因?),然而,它与TubB1的关系很疏远,就像它与其他生物中更保守的同源物的关系一样。