FUNCTIONAL-ANALYSIS OF NTF-1, A DEVELOPMENTALLY REGULATED DROSOPHILA TRANSCRIPTION FACTOR THAT BINDS NEURONAL CIS ELEMENTS
FUNCTIONAL-ANALYSIS OF NTF-1, A DEVELOPMENTALLY REGULATED DROSOPHILA TRANSCRIPTION FACTOR THAT BINDS NEURONAL CIS ELEMENTS
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DOI：
10.1101/gad.3.11.1677
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发表时间：
1989-11-01
影响因子：
10.5
通讯作者：
TJIAN, R
中科院分区：
文献类型：
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作者：
DYNLACHT, BD;ATTARDI, LD;TJIAN, R
为了研究在果蝇发育过程中调控基因表达的序列特异性转录因子，我们鉴定并纯化了一种新的DNA结合活性（NTF-1）。纯化的蛋白质由几种多肽组成，这些多肽选择性地结合至Ultrabithorax（Ubx）启动子的功能上重要的顺式控制元件以及多巴脱羧酶（Ddc）和fushi tarazu（Ftz）启动子/增强子区域的神经原性元件。纯化的NTF-1通过Ubx启动子的上游序列以结合位点依赖性方式在体外激活转录。一个cDNA克隆编码的开放阅读框架的NTF-1的分离，和推导的一级氨基酸序列的NTF-1包括谷氨酰胺丰富的区域，让人联想到转录激活结构域中发现的Sp1，但没有可识别的DNA结合结构域。NTF-1的表达在胚胎发育过程中受到时间调控。此外，原位杂交实验表明，NTF-1在胚胎中以空间受限的模式转录，在表皮和CNS中的细胞亚群中观察到最高水平的表达。NTF-1 cDNA在哺乳动物细胞中的表达产生了一种蛋白质，该蛋白质显示出与从果蝇胚胎分离的蛋白质的集合不可区分的DNA结合和转录活性。这些研究结果表明，NTF-1是一个家庭的成员的发育调节转录因子，可能参与指导表达的基因，如Ubx，Ddc，和ftz在神经元细胞。
In an effort to characterize sequence-specific transcription factors that regulate gene expression during Drosophila development, we identified and purified a novel DNA-binding activity (NTF-1). The purified protein consists of several polypeptides that bind selectively to a functionally important cis-control element of the Ultrabithorax (Ubx) promoter and to the neurogenic elements of both the dopa decarboxylase (Ddc) and fushi tarazu (ftz) promoter/enhancer regions. Purified NTF-1 activates transcription in vitro in a binding site-dependent manner through upstream sequences of the Ubx promoter. A cDNA clone encoding the open reading frame of NTF-1 was isolated, and the deduced primary amino acid sequence of NTF-1 includes a glutamine-rich region reminiscent of the transcriptional activation domains found in Sp1 but no recognizable DNA-binding domain. NTF-1 expression is temporally regulated during embryonic development. In addition, in situ hybridization experiments revealed that NTF-1 is transcribed in a spatially restricted pattern in the embryo, with the highest level of expression observed in the epidermis and a subset of cells in the CNS. Expression of the NTF-1 cDNA in mammalian cells yields a protein that displayed DNA-binding and transcriptional activities indistinguishable from that of the collection of proteins isolated from Drosophila embryos. These findings suggest that NTF-1 is a member of a family of developmentally regulated transcription factors that may be involved in directing expression of genes such as Ubx, Ddc, and ftz in neuronal cells.