Identification of the Doublesex protein binding sites that activate expression of lozenge in the female genital disc in Drosophila melanogaster
Identification of the Doublesex protein binding sites that activate expression of lozenge in the female genital disc in Drosophila melanogaster
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鉴定激活果蝇雌性生殖盘中菱形表达的 Doublesex 蛋白结合位点
DOI：
10.1016/j.mod.2017.01.001
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
and Suzuki MG.
中科院分区：
文献类型：
--
作者：
Wagamitsu S;Takase D;Aoki F;and Suzuki MG.
生殖器官的正常性分化对于有性生殖的动物物种是必不可少的。尽管已知doubletex（dsx）是多种昆虫生殖器性发育所必需的，但负责生殖器性分化的直接靶基因尚未确定。lz基因表达于雌性果蝇的生殖盘，是受精囊和附属腺发育所必需的。女性特异性的DSX亚型（DSXF）是激活女性生殖盘中lz表达所必需的。然而，DSXF是否直接激活雌性生殖盘中的转录仍然不清楚。在本研究中，我们在lz基因座内发现了两个与DSX结合基序高度同源的序列（lz-DBS 1和lz-DBS 2）。使用重组DSX DNA结合结构域（DSX-DBD）蛋白的竞争测定证实DSX-DBD蛋白以序列特异性方式与lz-DBS 1和lz-DBS 2结合，其亲和力低于与thebric-à-brac 1（bab 1）基因中已知DSX结合位点的亲和力。报告基因分析显示，含有lz-DBS 1和lz-DBS 2的2.5kb plz基因组片段以类似于雌性生殖盘中内源性lz表达的方式驱动报告基因（EGFP）表达。单独lz-DBS 1突变显著减少了EGFP表达区域的面积，而当两个位点都突变时，女性生殖盘中的EGFP表达被取消。这些结果表明DSX通过lz-DBS 1和lz-DBS 2直接激活生殖盘中的雌性特异性lz表达。
Normal sexual differentiation in the genital organs is essential for the animal species that use sexual reproduction. Although it is known thatdoublesex(dsx) is required for the sexual development of the genitalia in various insect species, the direct target genes responsible for the sexual differentiation of the genitalia have not been identified. Thelozenge(lz) gene is expressed in the female genital disc and is essential for developments of spermathecae and accessory glands inDrosophila melanogaster. The female-specific isoform of DSX (DSXF) is required for activatinglzexpression in the female genital disc. However, it still remains unclear whether the DSXF directly activates the transcription oflzin the female genital disc. In this study, we found two sequences (lz-DBS1 and lz-DBS2) withinlzlocus that showed high homoloty to the DSX binding motif identified previously. Competition assays using recombinant DSX DNA-binding domain (DSX-DBD) protein verified that the DSX-DBD protein bound to lz-DBS1 and lz-DBS2 in a sequence-specific manner with lower affinity than to the known DSX binding site in thebric-à-brac 1(bab1) gene. Reporter gene analyses revealed that a 2.5-kbplzgenomic fragment containing lz-DBS1 and lz-DBS2 drove reporter gene (EGFP) expression in a manner similar to endogenouslzexpression in the female genital disc. Mutations in lz-DBS1 alone significantly reduced the area of EGFP-expressing region, while EGFP expression in the female genital disc was abolished when both sites were mutated. These results demonstrated that DSX directly activates female-specificlzexpression in the genital disc through lz-DBS1 and lz-DBS2.
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影响因子：
10.5
作者：
Lingyang Zhu;J. Wilken;N. Phillips;U. Narendra;G. Chan;S. M. Stratton;S. Kent;M. Weiss
通讯作者：
Lingyang Zhu;J. Wilken;N. Phillips;U. Narendra;G. Chan;S. M. Stratton;S. Kent;M. Weiss
影响因子：
11.4
作者：
Kopp, Artyom
通讯作者：
Kopp, Artyom
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影响因子：
4
作者：
Wodarz,Andreas;Stewart,DanielB;Nelson,WJames;Nusse,Roel
通讯作者：
Nusse,Roel
影响因子：
10.5
作者：
COSCHIGANO, KT;WENSINK, PC
通讯作者：
WENSINK, PC
影响因子：
56.9
作者：
Lebestky, T;Chang, T;Banerjee, U
通讯作者：
Banerjee, U