Pearson L, Meagher R B
Westvaco Forest Science Laboratory, Summerville, SC 29484.
Plant Mol Biol. 1990 Apr;14(4):513-26. doi: 10.1007/BF00027497.
Plant actins are encoded by complex and highly divergent multigene families. Despite the general lack of intron conservation in animal, fungal and protist actin genes, evidence is presented which indicates that higher plant actin genes have an untranslated leader exon with structural similarity to that found in vertebrate actin genes. All functional higher plant actin genes sequenced to date contain a potential intron acceptor site in the 5' untranslated region 10 to 13 nucleotides upstream of the initiator ATG. A leader specific cDNA probe hybridized to sequences over 1.0 kbp upstream from the coding region confirming the presence of an upstream exon. Primer extension of mRNA with gene-specific oligonucleotides was used to analyze the 5' untranslated exon and leader intron from four divergent soybean actin genes, SAc3, 4, 6 and 7. The 5' ends of all four mRNAs are heterogeneous. The consensus promoter elements of the SAc7 actin promoter were identified. Gene specific primer extension sequencing of actin mRNAs indicated that splicing of the 5' leader intron occurred at the predicted acceptor site in SAc6 and SAc7. The SAc6 and SAc7 5' untranslated exons are small (88-111 nt) and the leader introns are relatively large (844-1496 nt). The presence of an intron within the 5' RNA leader and an intron which splits a glycine codon at position 152 in all plant actin genes and all vertebrate muscle actin genes suggests that these structures may have been conserved due to a functional role in actin expression. The 5' regions of these two soybean actin genes contain many unusual features including (CT) repeats and long stretches of pyrimidine-rich DNA. The possible roles of the upstream exon/intron and the C + T-rich regions are discussed.
植物肌动蛋白由复杂且高度分化的多基因家族编码。尽管动物、真菌和原生生物的肌动蛋白基因普遍缺乏内含子保守性,但有证据表明高等植物肌动蛋白基因有一个未翻译的前导外显子,其结构与脊椎动物肌动蛋白基因中的相似。迄今为止测序的所有功能性高等植物肌动蛋白基因在起始ATG上游10至13个核苷酸的5'非翻译区都含有一个潜在的内含子接受位点。一个前导特异性cDNA探针与编码区上游超过1.0 kbp的序列杂交,证实了上游外显子的存在。用基因特异性寡核苷酸对mRNA进行引物延伸,以分析四个不同的大豆肌动蛋白基因SAc3、4、6和7的5'非翻译外显子和前导内含子。所有四个mRNA的5'末端都是异质的。确定了SAc7肌动蛋白启动子的共有启动子元件。肌动蛋白mRNA的基因特异性引物延伸测序表明,SAc6和SAc7中5'前导内含子的剪接发生在预测的接受位点。SAc6和SAc7的5'非翻译外显子较小(88 - 111 nt),前导内含子相对较大(844 - 1496 nt)。所有植物肌动蛋白基因和所有脊椎动物肌肉肌动蛋白基因在5' RNA前导区内存在一个内含子,且在第152位的甘氨酸密码子处有一个内含子,这表明这些结构可能由于在肌动蛋白表达中的功能作用而得以保守。这两个大豆肌动蛋白基因的5'区域包含许多不寻常的特征,包括(CT)重复序列和长段富含嘧啶的DNA。讨论了上游外显子/内含子和富含C + T区域的可能作用。