Lucas Keira J, Zhao Bo, Roy Sourav, Gervaise Amanda L, Raikhel Alexander S
a Department of Entomology ; University of California, Riverside ; Riverside , CA USA.
b Graduate Program in Genetics; Genomics and Bioinformatics; University of California Riverside , Riverside , CA USA.
RNA Biol. 2015;12(12):1383-90. doi: 10.1080/15476286.2015.1101525.
Females of the hematophagous mosquito species require a vertebrate blood meal to supply amino acids and other nutrients necessary for egg development, serving as the driving force for the spread of many vector-borne diseases in humans. Blood digestion utilizes both early and late phase serine proteases (SPs) that are differentially regulated at the transcriptional and post-transcriptional level. To uncover the regulatory complexity of SPs in the female mosquito midgut, we investigated involvement of miRNAs in regulating the juvenile hormone (JH)-controlled chymotrypsin-like SP, JHA15. We identified regulatory regions complementary to the mosquito-specific miRNA, miR-1890, within the 3' UTR of JHA15 mRNA. The level of the JHA15 transcript is highest post eclosion and drastically declines post blood meal (PBM), exhibiting an opposite trend to miR-1890 that peaks at 24 h PBM. Depletion of miR-1890 results in defects in blood digestion, ovary development and egg deposition. JHA15 mRNA and protein levels are elevated in female mosquitoes with miR-1890 inhibition. JHA15 RNA interference in the miR-1890 depletion background alleviates miR-1890 depletion phenotypes. The miR-1890 gene is activated by the 20-hydroxyecdysone pathway that involves the ecdysone receptor and the early genes, E74B and Broad Z2. Our study suggests that miR-1890 controls JHA15 mRNA stability in a stage- and tissue- specific manner.
吸血蚊种的雌性需要摄取脊椎动物的血餐来提供卵子发育所需的氨基酸和其他营养物质,这是许多人类媒介传播疾病传播的驱动力。血液消化利用早期和晚期丝氨酸蛋白酶(SPs),它们在转录和转录后水平受到不同的调控。为了揭示雌性蚊子中肠中丝氨酸蛋白酶的调控复杂性,我们研究了微小RNA(miRNAs)在调控保幼激素(JH)控制的类胰凝乳蛋白酶SP,即JHA15中的作用。我们在JHA15 mRNA的3'非翻译区(UTR)内鉴定出与蚊子特异性miRNA miR-1890互补的调控区域。JHA15转录本的水平在羽化后最高,在血餐后(PBM)急剧下降,呈现出与miR-1890相反的趋势,miR-1890在血餐后24小时达到峰值。miR-1890的缺失导致血液消化、卵巢发育和卵子沉积出现缺陷。在miR-1890受到抑制的雌性蚊子中,JHA15 mRNA和蛋白质水平升高。在miR-1890缺失背景下对JHA15进行RNA干扰可减轻miR-1890缺失的表型。miR-1890基因由涉及蜕皮激素受体以及早期基因E74B和Broad Z2的20-羟基蜕皮激素途径激活。我们的研究表明,miR-1890以阶段和组织特异性的方式控制JHA15 mRNA的稳定性。