Ouyang C, Sprague K U
Department of Physics, University of Oregon, Eugene, OR 97403, USA.
Gene. 1998 Oct 23;221(2):207-13. doi: 10.1016/s0378-1119(98)00460-0.
The TATA-binding protein is a general transcription factor required by all three eukaryotic nuclear RNA polymerases. In order to study the function of this protein in the transcription of tRNA genes in the silkworm Bombyx mori, we have cloned TBP cDNA from a silkworm cDNA library. As in most other eukaryotes, TBP in silkworms is encoded by a single copy gene and contains a highly conserved C-terminal domain that includes a basic region and two direct repeats. In the less conserved N-terminal domain, silkworm TBP exhibits characteristics such as a glutamine-rich stretch and three imperfect Pro-Met-Thr-like repeats that are also found in Drosophila and human TBP. Silkworm TBP expressed in Escherichia coli and purified to apparent homogeneity binds the TATA element of the wild-type adenovirus major late promoter with nanomolar affinity.
TATA 结合蛋白是所有三种真核细胞核 RNA 聚合酶所需的通用转录因子。为了研究该蛋白在家蚕(Bombyx mori)tRNA 基因转录中的功能,我们从家蚕 cDNA 文库中克隆了 TBP cDNA。与大多数其他真核生物一样,家蚕中的 TBP 由单拷贝基因编码,并包含一个高度保守的 C 末端结构域,该结构域包括一个碱性区域和两个直接重复序列。在家蚕 TBP 不太保守的 N 末端结构域中,表现出富含谷氨酰胺的区域以及三个不完美的 Pro-Met-Thr 样重复序列等特征,这些特征在果蝇和人类 TBP 中也有发现。在大肠杆菌中表达并纯化至表观均一性的家蚕 TBP 以纳摩尔亲和力结合野生型腺病毒主要晚期启动子的 TATA 元件。