Ohbayashi T, Makino Y, Tamura T A
Department of Biology, Faculty of Science, Chiba University and CREST Japan Science and Technology Corporation, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan.
Nucleic Acids Res. 1999 Feb 1;27(3):750-5. doi: 10.1093/nar/27.3.750.
TATA-binding protein (TBP) is an essential factor for eukaryotic transcription. In this study, we demonstrated a mouse cDNA encoding a 21 kDa TBP-like protein (TLP). The TLP ORF, carrying 186 amino acids, covered the entire 180 amino acids of the C-terminal conserved domain of mouse TBP with 39% identity and 76% similarity. Northern blot analysis demonstrated that TLP mRNAs were expressed in various mammalian tissues ubiquitously and that their distribution pattern was analogous to that of TBP. By using anti-TLP antibody, we demonstrated the existence of TLP proteins in various mammalian cells and tissues. The Drosophila TBP-related factor (TRF) is a neurogenesis-related transcription factor that binds to the TATA-box and activates transcription. TLP did not bind to the TATA-box nor direct transcription initiation. Multiple amino acids critical for TBP function were deleted or substituted in TLP, while amino acids in Drosophila TRF much resembled those in TBP. Similarity between Drosophila TRF and mouse TLP was considerably lower (alignment score 35) than that between Drosophila TBP and mouse TBP (alignment score 88). Identity of nucleotide sequences between mouse and putative human TLPs (94%) was higher than that between TBPs (91%) in these two animals. Expression of TLP was nearly constant throughout the P19 differentiation process. Accordingly, we suggest that, even if higher eukaryotes generally contain multiple tbp -related genes, TLP is not a bona fide mammalian counterpart of Drosophila TRF.
TATA 结合蛋白(TBP)是真核生物转录的必需因子。在本研究中,我们展示了一个编码 21 kDa TBP 样蛋白(TLP)的小鼠 cDNA。TLP 的开放阅读框(ORF)包含 186 个氨基酸,覆盖了小鼠 TBP C 端保守结构域的全部 180 个氨基酸,同源性为 39%,相似性为 76%。Northern 印迹分析表明,TLP mRNA 在各种哺乳动物组织中普遍表达,其分布模式与 TBP 类似。通过使用抗 TLP 抗体,我们证明了 TLP 蛋白在各种哺乳动物细胞和组织中的存在。果蝇 TBP 相关因子(TRF)是一种与神经发生相关的转录因子,它与 TATA 框结合并激活转录。TLP 不与 TATA 框结合,也不指导转录起始。TBP 功能所必需的多个氨基酸在 TLP 中被缺失或替换,而果蝇 TRF 中的氨基酸与 TBP 中的非常相似。果蝇 TRF 与小鼠 TLP 之间的相似性(比对得分 35)远低于果蝇 TBP 与小鼠 TBP 之间的相似性(比对得分 88)。在这两种动物中,小鼠与推定的人类 TLP 之间核苷酸序列的同源性(94%)高于 TBP 之间的同源性(91%)。在整个 P19 分化过程中,TLP 的表达几乎保持恒定。因此,我们认为,即使高等真核生物通常含有多个与 tbp 相关的基因,但 TLP 并非果蝇 TRF 在哺乳动物中的真正对应物。