2006-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/658847摘要：在有絲分裂過程中染色體分離受到嚴密的監控，當染色體分離有缺失時，會導致多倍體而造成細胞和生物體的死亡或細胞轉型和腫瘤的產生。線蟲TLK-1激?對於有絲分裂染色體的濃縮和分離是必要的，tlk-1 RNAi的胚胎具非整數倍體染色 體，我們初步的研究顯示TLK-1藉著將HCP-3帶到有絲分裂DNA的極端面 (poleward face)，來調控有絲分裂染色體的結構，進而影響有絲分裂染色體的分離，這個功能是之前尚未被發現的，然而TLK-1如何控制DNA濃縮仍未明瞭。在這個計劃中，我們將使用質譜儀基礎的蛋白質體學方式，來搜尋與TLK-1結合的蛋白質，並利用RNAi來研究這些蛋白質的功能，再進一步結合生化分析，以了解TLK-1調控DNA濃縮和分離的機制。如果時間和預算允許下，我們會利用先前引入牛痘病毒到線蟲中所建立的PEG-mediated方式，更深入研究TLK-1及其結合蛋白在包裹病毒基因組中的可能角色，這將會幫助我們了解病毒粒子成熟的最後階段中病毒和宿主間的可能交互作用。另外有趣的是我們初步研究顯示人類Tlk2可以部份援助tlk-1(RNAi)胚胎的致死率，表示tlk-1的功能在演化上<br> Abstract: Chromosome segregation during mitosis is under strict control. Defects in chromosome segregation leading to changes in ploidy may cause cellular and organismal death or in cellular transformation and tumorigenesis. C. elegans TLK-1 kinase is important for mitotic chromosome condensation and segregation. The tlk-1 (RNAi) embryos were aneuploid. Our preliminary data showed that TLK-1 likely promotes mitotic chromosome architecture by localizing HCP-3 to the poleward face of mitotic DNA, a role not described previously, and, as a consequence, controls mitotic chromosome segregation. However, how TLK-1 controls DNA condensation is still unclear. In this project we aim to use mass-spectrometry-based proteomics approach to identify TLK-1 interacting proteins. We will then generate and characterize deletion alleles for the corresponding genes. Such studies will reveal molecules that act together with TLK-1. Further biochemical analysis of their interactions will help elucidate the mechanisms of TLK-1-mediated DNA condensation and segregation. If permitted by time and budget, we will further investigate the potential effect of TLK-1 and its interactors in packaging viral genome using the PEG-mediated method we previously established to introduce vaccinia virus into C. elegans. This will help us understand the potential interactions between virus and host at the late stage of virion maturation. Interestingly, our preliminary data showed that human Tlk2 can partially rescue the lethality of tlk-1(RNAi) embryos, suggesting the function of tlk-1 is conserved in evolution. We believe that our studies of TLK-1 interactors will contribute to our knowledge in DNA condensation and segregation, a conserved and important cell cycle event, in C. elegans and humans, and even may reveal a role in virus-host interaction at the level of viral genome packaging.