May G S, Tsang M L, Smith H, Fidel S, Morris N R
Department of Pharmacology, UMDNJ Robert Wood Johnson Medical School, Piscataway 08854.
Gene. 1987;55(2-3):231-43. doi: 10.1016/0378-1119(87)90283-6.
Aspergillus nidulans has two beta-tubulin genes: benA, which is involved in both vegetative growth and asexual sporulation, and tubC, which is involved mainly in asexual sporulation. Both genes have now been cloned and sequenced. benA encodes a polypeptide of 447 amino acids (aa) and tubC encodes one of 449 aa. The two polypeptides differ by 78 aa residues but the net charge for the two proteins remains the same. The divergence between the amino acid sequences of the Aspergillus beta-tubulins is greater than that for any other two beta-tubulins yet described in the same organism. The benA gene has eight introns and the tubC gene has five, all of which correspond in amino acid position to introns in benA. The positions of some of these introns are conserved in other beta-tubulin genes. The 5'-splice site, internal, and 3'-splice site consensus sequences are similar to those found in other fungal introns. The transcriptional start points for each gene have been determined using primer extension and/or S1 nuclease mapping. Neither the benA gene nor the tubC gene contains a TATA sequence in its 5'-flanking region. The tubC gene has two repeated upstream sequences which are not found in benA. The sites of polyadenylation have been determined for each gene using S1 nuclease mapping. Neither gene contains a polyadenylation signal, AATAAA, typical of other eukaryotic genes.
构巢曲霉有两个β-微管蛋白基因:benA,它参与营养生长和无性孢子形成;tubC,它主要参与无性孢子形成。现在这两个基因都已被克隆和测序。benA编码一个由447个氨基酸(aa)组成的多肽,tubC编码一个由449个aa组成的多肽。这两个多肽相差78个aa残基,但这两种蛋白质的净电荷保持相同。构巢曲霉β-微管蛋白的氨基酸序列之间的差异大于同一生物体中迄今描述的任何其他两个β-微管蛋白之间的差异。benA基因有8个内含子,tubC基因有5个内含子,所有这些内含子在氨基酸位置上都与benA中的内含子相对应。其中一些内含子的位置在其他β-微管蛋白基因中是保守的。5'-剪接位点、内部和3'-剪接位点的共有序列与其他真菌内含子中的序列相似。使用引物延伸和/或S1核酸酶作图确定了每个基因的转录起始点。benA基因和tubC基因在其5'-侧翼区域都不包含TATA序列。tubC基因有两个在benA中未发现的上游重复序列。使用S1核酸酶作图确定了每个基因的聚腺苷酸化位点。这两个基因都不包含其他真核基因典型的聚腺苷酸化信号AATAAA。