Molecular cloning and identification of the transcriptional regulatory domain of the goat neurokinin B gene TAC3.
Molecular cloning and identification of the transcriptional regulatory domain of the goat neurokinin B gene TAC3.
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分子克隆和识别山羊神经蛋白B基因TAC3的转录调节结构域。
DOI：
10.1262/jrd.2013-037
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发表时间：
2013-10
期刊：
影响因子：
--
通讯作者：
Ohkura S
中科院分区：
文献类型：
--
作者：
Suetomi Y;Matsuda F;Uenoyama Y;Maeda K;Tsukamura H;Ohkura S
由TAC 3编码的神经激肽B（NKB）被认为是促性腺激素释放激素脉冲释放的重要加速剂。本研究旨在阐明山羊TAC 3的转录调控机制。首先，我们确定山羊下丘脑TAC 3的全长mRNA序列为820 B，包括381 B编码区，假定的转录起始位点位于起始密码子上游143-B。由preproNKB产生的NKB的推导的氨基酸序列在山羊、牛和人中是完全保守的。其次，我们克隆了山羊TAC 3的5 '端上游区域，该区域位于翻译起始位点3400 B处，与牛TAC 3的同源性为89%。我们使用该山羊TAC 3 5 '-上游区进行荧光素酶测定。我们创建了一个荧光素酶报告载体，该载体含有山羊TAC 3的-2706、-1837、-834、-335或-197至+166 bp（假定的转录起始位点被指定为+1）的DNA构建体，并将其瞬时转染到小鼠下丘脑衍生的N7细胞和人神经母细胞瘤衍生的SK-N-AS细胞中。荧光素酶活性随着5 '端上游区域的缺失而逐渐增强，表明转录抑制区位于-2706 ~-336 bp之间，核心启动子位于-197 bp下游。Escherichia coli处理没有导致任何构建体的荧光素酶活性的显著抑制，表明存在调节山羊TAC 3转录的其他因子。
Neurokinin B (NKB), encoded by TAC3, is thought to be an important accelerator of pulsatile gonadotropin-releasing hormone release. This study aimed to clarify the transcriptional regulatory mechanism of goat TAC3. First, we determined the full-length mRNA sequence of goat TAC3 from the hypothalamus to be 820 b, including a 381 b coding region, with the putative transcription start site located 143-b upstream of the start codon. The deduced amino acid sequence of NKB, which is produced from preproNKB, was completely conserved among goat, cattle, and human. Next, we cloned 5’-upstream region of goat TAC3 up to 3400 b from the translation initiation site, and this region was highly homologous with cattle TAC3 (89%). We used this goat TAC3 5’-upstream region to perform luciferase assays. We created a luciferase reporter vector containing DNA constructs from –2706, –1837, –834, –335, or –197 to +166 bp (the putative transcription start site was designated as +1) of goat TAC3 and these were transiently transfected into mouse hypothalamus-derived N7 cells and human neuroblastoma-derived SK-N-AS cells. The luciferase activity gradually increased with the deletion of the 5’-upstream region, suggesting that the transcriptional suppressive region is located between –2706 and –336 bp and that the core promoter exists downstream of –197 bp. Estradiol treatment did not lead to significant suppression of luciferase activity of any constructs, suggesting the existence of other factor(s) that regulate goat TAC3 transcription.
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影响因子：
1.8
作者：
Noritake, Ken-ichi;Matsuoka, Toshiki;Tsukamura, Hiroko
通讯作者：
Tsukamura, Hiroko
影响因子：
56.9
作者：
BELCHETZ, PE;PLANT, TM;KNOBIL, E
通讯作者：
KNOBIL, E
影响因子：
5.3
作者：
Otto, Stefanie J.;McCorkle, Sean R.;Mandel, Gail
通讯作者：
Mandel, Gail
影响因子：
4.8
作者：
DIERSCHK.DJ;BHATTACH.AN;KNOBIL, E
通讯作者：
KNOBIL, E
影响因子：
56.9
作者：
KNOBIL, E;PLANT, TM;MARSHALL, G
通讯作者：
MARSHALL, G