Investigating the molecular functions and mechanisms of RNA-binding proteins crucial for gametogenesis

在试图怀孕的夫妇中，10%-15%的人会受到不孕不育的影响，这可能是由 配子发生受损的。RNA结合蛋白对转录后基因的有效调控 对正常的配子发生至关重要，限制性商业惯例的突变破坏了这种调节并导致 人类的不孕不育。例如，在无精子症(DAZ)家族中缺失的限制性商业惯例对于 人类生育能力，因为在精子发生期间，它们通过以下方式调节mRNA子集的翻译 与它们的3‘UTRs结合；它们的分子作用机制只有在它们的 对果蝇直系同源基因进行了鉴定和遗传学研究。然而，所有的RBP曲目都是 配子发生所必需的物质目前还不清楚。要充分理解复杂的转录后转录 在配子发生过程中，识别对配子发生至关重要的新限制性商业惯例是至关重要的。 并阐明它们的分子功能和作用机制。 我们最近发现了三个以前没有特征的RBP(MARF1、Tanenashi和 CG44249)，它们对果蝇的生育能力是必不可少的。我们的数据表明MARF1，也就是 在卵母细胞中表达，是女性生育所必需的，在 卵子发生。Tanenashi在精母细胞中表达，是男性生育所必需的，发挥作用 巨碱基对男性生育因子基因表达和剪接的促进作用 精子发生过程中Y染色体上DNA序列高度重复的内含子。CG44249， 它在卵巢中表达，是女性生育所必需的，可能在剪接调节中发挥作用 在卵子发生过程中。我们还发现了第四个RBP(Loqs-PB)，它对女性生育能力很重要 生殖系干细胞在卵巢中的维持，调节卵巢中microRNAs的长度。这些 从苍蝇到人类，有四种限制性商业惯例是保守的。 利用果蝇，我现在提议进一步研究这四种限制性商业惯例，并确定它们的 配子发生中精确的分子功能和作用机制。通过研究突变苍蝇 在体内和体外的蛋白质中，我们将检验我们的假设：(I)这些限制性商业惯例各自结合特定的靶点 通过识别特定的RNA序列和/或结构基序来识别RNA，以及(Ii)从而促进或抑制 在特定的分子步骤上处理结合的RNA，允许高度调控的时空 靶基因在配子发生过程中的表达。 这些研究将促进我们对后遗症分子机制的基础知识。 基因表达的转录调节，特别是通过调节信使核糖核酸Poly-A尾长， 在配子发生过程中的mRNA剪接和microRNA长度，提供了改进的可能性 人类不孕不育的诊断和治疗。

Among couples trying to conceive, 10-15% will be affected by infertility, which can be caused by impaired gametogenesis. Effective post-transcriptional gene regulation by RNA-binding proteins (RBPs) is crucial for normal gametogenesis, with mutations in RBPs impairing such regulation and causing infertility in humans. For example, the Deleted in Azoospermia (DAZ) family of RBPs are essential for human fertility, because during spermatogenesis they regulate translation of a subset of mRNAs by binding to their 3′ UTRs; their molecular mechanism of action was understood only after their Drosophila ortholog was identified and studied genetically. However, the full repertoire of RBPs that are essential for gametogenesis is currently unknown. To fully understand the complex post-transcriptional gene regulation during gametogenesis, it is critical to identify novel RBPs that are important for these processes and elucidate their molecular functions and mechanisms of action. We have recently identified three previously uncharacterized RBPs (MARF1, Tanenashi, and CG44249) that are essential for fertility in Drosophila. Our data suggest that MARF1, which is expressed in oocytes and is required for female fertility, regulates mRNA poly-A tail length during oogenesis. Tanenashi, which is expressed in spermatocytes and is required for male fertility, plays roles in promoting the expression and splicing of male fertility factor genes containing mega-base-sized introns with highly repetitive DNA sequence on the Y chromosome during spermatogenesis. CG44249, which is expressed in ovaries and is required for female fertility, likely plays a role in splicing regulation during oogenesis. We also showed that a fourth RBP (Loqs-PB), which is important for female fertility and germline stem cell maintenance in ovaries, regulates the length of microRNAs in ovaries. These four RBPs are conserved from flies to humans. Using Drosophila, I now propose to further investigate these four RBPs, and determine their precise molecular functions and mechanisms of action in gametogenesis. By investigating mutant flies in vivo and the proteins in vitro, we will test our hypothesis that: (i) these RBPs each bind specific target RNAs by recognizing specific RNA sequence and/or structure motifs, and (ii) thereby promote or inhibit the processing of bound RNAs at specific molecular steps, allowing highly regulated spatiotemporal expression of targeted genes during gametogenesis. These studies will advance our fundamental knowledge of the molecular mechanisms of post- transcriptional regulation of gene expression, especially by regulating the mRNA poly-A tail length, mRNA splicing, and microRNA length, during gametogenesis, providing potential for improved diagnosis and treatment of human infertility.