DNA replication and cytokinesis

描述(申请人提供)：起源识别复合体(ORC)，异构体六亚单位蛋白质复合体，对DNA复制是必不可少的。ORC的功能也超越了DNA复制。在我们的初步研究中，我们发现果蝇最小的ORC亚基ORC6对于DNA结合和DNA复制是重要的，但也通过与Septin蛋白Pnut的相互作用而在细胞质分裂中发挥作用。基于这些观察结果，我们推测ORC6是一种多功能蛋白质。ORC6可能参与协调细胞分裂周期的许多过程的信号通路，如染色体复制和胞质分裂。我们推测，ORC6与转录因子TFIIB在结构上具有同源性，对于ORC靶向果蝇细胞的起源和形成复制前复合体是必不可少的。ORC6在胞质分裂中的作用是通过调节Septin复合体的活性来实现的。为了验证这些假设，我们计划在分子、细胞和遗传水平上分析ORC6的功能和活性，以及它在细胞周期中的作用。具体地说，我们将1)确定ORC6的三维结构；2)确定ORC6在DNA结合和DNA复制中的功能；以及3)阐明ORC6在调节间隔蛋白复合体功能中的作用。由于ORC在进化上是保守的，我们对其在果蝇中的功能的了解将对我们理解ORC在其他真核生物中的功能具有重要意义。更好地了解ORC在果蝇中的功能将为深入了解ORC在哺乳动物中的行为提供线索，并为其错误调控如何导致癌症提供线索。

DESCRIPTION (provided by applicant): The Origin Recognition Complex (ORC), heteromeric six-subunit protein complex, is essential for DNA replication. ORC's functions also extend beyond DNA replication. In our preliminary studies, we found that the Drosophila smallest ORC subunit, Orc6, is important for DNA binding and DNA replication but also has a function in cytokinesis through the interaction with septin protein Pnut. Based on these observations we hypothesize that Orc6 is a multifunctional protein. Orc6 might be involved in a signaling pathway that coordinates many processes of the cell division cycle, such as chromosome duplication and cytokinesis. We hypothesize that Orc6, which bears a structural homology with transcription factor, TFIIB, is essential for targeting of ORC to the origins in Drosophila cells and for the formation of pre- replicative complex. The function of Orc6 in cytokinesis is achieved through the regulation of septin complex activities. To test these hypotheses we plan to analyze at the molecular, cellular and genetic levels the functions and activities of Orc6 and its role in cell cycle. Specifically we will 1) determine the three-dimensional structure of Orc6; 2) detertmine functions of Orc6 in DNA binding and DNA replication; and 3) elucidate the role of Orc6 in regulation of septin complex functions. Because ORC is conserved in evolution, what we learn about its functions in Drosophila will be valuable to our understanding of ORC's functions in other eukaryotes. A better understanding of how ORC functions in Drosophila will provide insights into ORC's behavior in mammals and provide clues as to how its misregulation leads to cancer.