Molecular characterization of two novel milk proteins in the tsetse fly (Glossina morsitans morsitans).
Molecular characterization of two novel milk proteins in the tsetse fly (Glossina morsitans morsitans).
复制标题
采采蝇（Glossina morsitans morsitans）中两种新型乳蛋白的分子特征。
DOI：
10.1111/j.1365-2583.2009.00987.x
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发表时间：
2010
影响因子：
2.6
通讯作者：
Aksoy,S
中科院分区：
文献类型：
--
作者：
Yang,G;Attardo,GM;Lohs,C;Aksoy,S
目的：乳蛋白是采采蝇胎生生殖的重要组成部分。乳蛋白在乳腺组织中合成并分泌，占子宫内幼虫营养所需分泌物的50%。方法：利用生物信息学方法从采采蝇组织特异性cDNA文库中筛选出两个未知分泌蛋白（GmmMGP 2和GmmMGP 3）。进行RT-PCR分析以验证其乳腺/脂肪体特异性表达谱。通过北方印迹分析和荧光原位杂交对这些蛋白质的发育和组织特异性表达进行了详细的表征。结果：预测的gmmmgp 2和gmmmgp 3蛋白分子量均为22 kD左右，含有较高比例的疏水氨基酸和潜在的磷酸化位点。这两种基因的表达是乳腺分泌细胞的组织特异性。这两个基因的转录丰度增加的过程中，子宫内幼虫的发展和平行的ofgmmmgp，一个很好的特点牛奶蛋白基因被认为是主要的牛奶蛋白。RNA干扰处理后果蝇的表型分析显示，gmmmgp 2基因敲除果蝇的生殖力有显著影响，而gmmmgp 3基因敲除果蝇的生殖力无显著影响。敲低gmmmgp 2导致排卵中断和随之而来的卵母细胞积累和degradation.Gmmmgp2敲低也有显着的影响苍蝇mortals.Conclusions：这项工作确定了两个新的基因，蛋白质的功能，在子宫内幼虫在采采蝇。这些蛋白质可能是从母体提供给幼虫的乳汁分泌物的营养成分。敲除gmmmgp 2的表型数据表明，鉴于在敲除果蝇中观察到的排卵缺陷，这种蛋白质也可能具有调节功能。这些基因的进一步分析将是重要的（与其他乳蛋白），直接乳腺/妊娠特异性基因表达的转录调控机制的鉴定。
Purpose: Milk proteins are an essential component of viviparous reproduction in the tsetse fly. Milk proteins are synthesized in and secreted from the milk gland tissue and constitute 50% of the secretions from which the intrauterine larva derives its nourishment. To understand milk protein function and regulation during viviparous reproduction, milk proteins need to be identified and characterized.Methods: Two putative unknown secretory proteins (GmmMGP2 and GmmMGP3) were selected by bioinformatic analysis of tissue specific tsetse cDNA libraries. RT‐PCR analysis was performed to verify their milk gland/fat body specific expression profile. Detailed characterization of developmental and tissue specific expression of these proteins was performed by northern blot analysis and fluorescentin situhybridization. Functional analysis of the milk gland proteins during the tsetse gonotrophic cycle was performed using RNA interference (RNAi).Results: The predicted proteins fromgmmmgp2andgmmmgp3are small ∼22 kD and contain a high proportion of hydrophobic amino acids and potential phosphorylation sites. Expression of both genes is tissue specific to the secretory cells of the milk gland. Transcript abundance for both genes increases over the course of intrauterine larval development and parallels that ofgmmmgp, a well characterized milk protein gene considered to be the major milk protein. Phenotypic analysis of flies after RNA interference treatment revealed a significant effect upon fecundity in thegmmmgp2knockdown flies, but not thegmmmgp3flies. Knockdown ofgmmmgp2resulted in disruption of ovulation and consequent oocyte accumulation and degradation.Gmmmgp2knockdown also had a significant impact on fly mortality.Conclusions: This work identifies two novel genes, the proteins of which appear to function in response to intrauterine larvigenesis in tsetse. These proteins may be nutritional components of the milk secretions provided to the larva from the mother. Phenotypic data from knockdown ofgmmmgp2suggests that this protein may also have a regulatory function given the defect in ovulation observed in knockdown flies. Further analysis of these genes will be important (in conjunction with other milk proteins) for identification of transcriptional regulation mechanisms that direct milk gland/pregnancy specific gene expression.