MicroRNA-275 targets sarco/endoplasmic reticulum Ca2+ adenosine triphosphatase (SERCA) to control key functions in the mosquito gut.
MicroRNA-275 targets sarco/endoplasmic reticulum Ca2+ adenosine triphosphatase (SERCA) to control key functions in the mosquito gut.
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DOI：
10.1371/journal.pgen.1006943
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发表时间：
2017-08
期刊：
影响因子：
4.5
通讯作者：
Raikhel AS
中科院分区：
文献类型：
--
作者：
Zhao B;Lucas KJ;Saha TT;Ha J;Ling L;Kokoza VA;Roy S;Raikhel AS
黄热病蚊子埃及伊蚊是虫媒病毒的主要媒介，造成许多毁灭性的人类疾病，如登革热和黄热病、基孔肯雅热和寨卡病毒。雌蚊需要脊椎动物的血液来产卵，而反复取血的循环与病原体传播密切相关。蚊子的后中肠（肠道）参与血液消化，也是病原体的入口。因此，蚊子肠道是一个重要的研究组织。miRNA aae-miR-275 （miR-275）已被证明是雌性蚊子正常血液消化所必需的；然而，其作用机制尚不清楚。在这里，我们证明miR-275直接靶向并积极调节sarco/内质网Ca2+腺苷三磷酸酶，这涉及Ca2+从细胞质到sarco/内质网的主动运输。我们利用肠道特异性酵母转录激活蛋白Gal4/上游激活序列（Gal4/UAS）系统和miRNA Tough Decoy技术的组合来消耗转基因蚊子肠道内源性miR-275水平。血餐后肠道特异性miR-275的减少降低了SERCA mRNA和消化酶晚胰蛋白酶的蛋白质水平。它还导致肠道微生物群的显著减少。此外，miR-275和SERCA的降低与Notch信号通路的缺陷和肠肌动蛋白细胞骨架的组装相关。miR-275沉默引起的不良表型可以通过注射miR-275模拟物来修复。因此，我们发现miR-275直接靶向SERCA，维持其水平对蚊子的多种肠道功能至关重要。雌性蚊子传播许多毁灭性的人类疾病。蚊子肠道除了具有血液消化的主要功能外，还是病原体在蚊子载体中定植的入口。保守的microRNA miR-275被证明是血液消化和卵子发育所必需的。在这项研究中，我们研究了miR-275对蚊子肠道功能调节的作用靶点。我们利用转基因Tough诱饵RNA方法在埃及伊蚊雌蚊肠道中实现了miR-275的时空抑制。此外，我们发现miR-275靶向sarco/内质网Ca2+-腺苷三磷酸酶（SERCA），影响许多肠道功能，包括血液消化、消化蛋白酶的产生和肠肌动蛋白细胞骨架的组装。SERCA对维持Ca2+稳态至关重要，在人类中，SERCA的紊乱会导致心脏肥厚、心力衰竭和癌症。因此，miRNA miR-275靶向SERCA的发现不仅有助于了解蚊子肠道调控，而且显著增加了对这一关键分子调控机制的一般理解。
The yellow fever mosquito Aedes aegypti is the major vector of arboviruses, causing numerous devastating human diseases, such as dengue and yellow fevers, Chikungunya and Zika. Female mosquitoes need vertebrate blood for egg development, and repeated cycles of blood feeding are tightly linked to pathogen transmission. The mosquito’s posterior midgut (gut) is involved in blood digestion and also serves as an entry point for pathogens. Thus, the mosquito gut is an important tissue to investigate. The miRNA aae-miR-275 (miR-275) has been shown to be required for normal blood digestion in the female mosquito; however, the mechanism of its action has remained unknown. Here, we demonstrate that miR-275 directly targets and positively regulates sarco/endoplasmic reticulum Ca2+ adenosine triphosphatase, which is implicated in active transport of Ca2+ from the cytosol to the sarco/endoplasmic reticulum. We utilized a combination of the gut-specific yeast transcription activator protein Gal4/upstream activating sequence (Gal4/UAS) system and miRNA Tough Decoy technology to deplete the endogenous level of miR-275 in guts of transgenic mosquitoes. This gut-specific reduction of miR-275 post blood meal decreased SERCA mRNA and protein levels of the digestive enzyme late trypsin. It also resulted in a significant reduction of gut microbiota. Moreover, the decrease of miR-275 and SERCA correlated with defects in the Notch signaling pathway and assembly of the gut actin cytoskeleton. The adverse phenotypes caused by miR-275 silencing were rescued by injections of miR-275 mimic. Thus, we have discovered that miR-275 directly targets SERCA, and the maintenance of its level is critical for multiple gut functions in mosquitoes. Female mosquitoes transmit numerous devastating human diseases. The mosquito gut, in addition to its primary function as a site of blood digestion, represents the entry point for pathogen colonization in mosquito vectors. The conserved microRNA, miR-275, was shown to be required for blood digestion and egg development. In this study, we investigated the target of miR-275 contributing to the regulation of mosquito gut functions. We achieved spatiotemporal suppression of miR-275 using a transgenic Tough Decoy RNA approach in the A. aegypti female mosquito gut. Furthermore, we have uncovered that miR-275 targets sarco/endoplasmic reticulum Ca2+- adenosine triphosphatase (SERCA), affecting numerous gut functions including blood digestion, production of digestive proteases, and assembly of the gut actin cytoskeleton. SERCA is essential for maintenance of Ca2+ homeostasis, and its disturbance, in humans, leads to cardiac hypertrophy, heart failure and cancers. Therefore, the finding that the miRNA miR-275 targets SERCA not only contributes to the knowledge of mosquito gut regulation but also significantly adds to the general understanding of mechanisms governing this critical molecule.
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影响因子：
37.8
作者：
Gurha P;Abreu-Goodger C;Wang T;Ramirez MO;Drumond AL;van Dongen S;Chen Y;Bartonicek N;Enright AJ;Lee B;Kelm RJ Jr;Reddy AK;Taffet GE;Bradley A;Wehrens XH;Entman ML;Rodriguez A
通讯作者：
Rodriguez A
影响因子：
8.8
作者：
Clark RI;Salazar A;Yamada R;Fitz-Gibbon S;Morselli M;Alcaraz J;Rana A;Rera M;Pellegrini M;Ja WW;Walker DW
通讯作者：
Walker DW
蚊子特异性 microRNA-1174 靶向丝氨酸羟甲基转移酶以控制肠道关键功能
DOI：
10.1073/pnas.1416278111
发表时间：
2014-10-07
影响因子：
11.1
作者：
Liu, Shiping;Lucas, Keira J.;Raikhel, Alexander S.
通讯作者：
Raikhel, Alexander S.
影响因子：
4
作者：
Berridge, MJ
通讯作者：
Berridge, MJ
影响因子：
14.9
作者：
Haraguchi T;Ozaki Y;Iba H
通讯作者：
Iba H