The Role of a Novel PIWI-Interacting Protein in Spermatogenesis

描述(申请人提供)：遗传物质从一代传给下一代，即从父母传给他们的孩子，是一个关键的生物学过程。然而，人们对这一努力是如何实现的只有部分了解。我们之前已经发现一种称为Piwi蛋白的蛋白质家族在这一过程中是必不可少的。为了更详细地了解Piwi是如何工作的，我们已经确定了与其相互作用的其他蛋白质。在苍蝇中，Piwi与一种名为PAPI的蛋白质相互作用；这种相互作用在进化上是保守的，因为在老鼠和人类中，Piwi和PAPI蛋白也相互作用。在果蝇中，PAPI是胚胎正常发育和形成专门细胞(称为生殖系)所必需的，这些细胞将遗传信息传递给后代。然而，对于它在老鼠和人类等脊椎动物中如何发挥作用，我们一无所知。由于PAPI对果蝇胚胎的正常形成及其传递遗传信息的能力是必需的，而且它与Piwi蛋白的相互作用是高度保守的，我们假设这种相互作用对哺乳动物PAPI蛋白的功能是重要的。在这里，我们建议进行实验，以检验这一假设，并确定小鼠PAPI(MAPI)在生殖系调控中的作用。具体地说，我们建议1)研究MAPI在生殖系发育和成熟性腺中的表达模式，2)确定MAPI在出生后精子发生过程中的功能，以及3)鉴定MAPI蛋白和RNA复合体。 与公共卫生相关：有性繁殖生物，如老鼠和人类，需要将他们的遗传信息代代相传。这必须准确地完成，因为错误可能会导致后代出现严重的健康问题，并且需要体内高度专门化的组织来做到这一点。这项计划中的实验将确定一种名为MAPI的蛋白质如何参与这一过程。

DESCRIPTION (provided by applicant): The passing of genetic material from one generation to the next, i.e. from parents to their children, is a key biological process. How this endeavor is accomplished, however, is only partially understood. We have previously found a family of proteins called the Piwi proteins to be essential for this process. To understand in more detail how Piwi works, we have identified additional proteins which interact with it. In flies, Piwi interacts with a protein called Papi; this interaction is evolutionarily conserved, as Piwi and Papi proteins in mouse and human also interact. In flies, Papi is required for proper development of the embryo and for formation of specialized cells (called germline) which pass genetic information to future generations. However, nothing is known about how it may function in vertebrates such as mice and humans. Because Papi is required for proper formation of fly embryos and their ability to transmit genetic information, and because its interaction with Piwi proteins are highly conserved, we hypothesize that this interaction is important for the function of mammalian Papi proteins. Here, we propose experiments which will test this hypothesis and determine the role of Mouse Papi (Mapi) in regulation of the germline. Specifically, we propose to 1) examine the expression patterns of Mapi during germline development and in mature gonads, 2) determine the function of Mapi during post-natal spermatogenesis, and 3) characterize Mapi protein and RNA complexes. PUBLIC HEALTH RELEVANCE: Sexually reproducing organisms, such as mice and humans, need to pass their genetic information from one generation to the next. This must be accomplished accurately, as mistakes can lead to severe health issues in the offspring, and requires highly specialized tissue in the body to do so. The experiments in this proposal will define how one protein, Mapi, contributes to this process.