Functional cloning of the human RHAMM in complementation of cytokinesis-checkpoint defect in fission yeast

• Main Authors: Ko-Chia Ho, 何可嘉
• Other Authors: Jin-Yuan Su
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/11247479376264964316

碩士 === 國立陽明大學 === 生物化學研究所 === 91 === Cell-cycle checkpoints are known as surveillance mechanisms designed to ensure accurate transmission of genetic information during the progression of cell division. Spatial and temporal controls of cytokinesis ensure the coordination of mitosis and cytokinesis. However, the molecular basis for the cytokinesis checkpoint is still unclear. The fission yeast Schizosaccharomyces pombe has been established in recent years as a powerful model system for studying eukaryotic cytokinesis. In the fission yeast, a signal transduction pathway termed septation-initiation network (SIN) functions to regulate initiation of cytokinesis at the end of mitosis and to coordinate mitosis and cytokinesis. The fission yeast cdc16 gene, which belongs to components of SIN, encodes a GTPase-activating protein (GAP) and functions as a negative regulator of SIN pathway to stop septation. In order to understand the cytokinesis checkpoint mechanism from a different aspect, I have used a fission yeast temperature-sensitive mutant, cdc16-116, as a model system to search for human genes that can functionally complement cdc16-116 at the restricted temperature. I demonstrated that a truncated form of human RHAMM can functionally complement the cdc16-116 mutant and partially rescue its viability. To further explore the mechanism of RHAMM in the fission yeast cdc16-116 mutant, I performed yeast two-hybrid screening to search for RHAMM-interacting proteins and identified three yeast proteins including Clr3p, Cdc10p and nucleoporin that specifically interact with RHAMM. RHAMM presumably acts by interacting with these proteins to overcome the cytokinesis defect in fission yeast.