Genome-Wide Deficiency Screen for the Genomic Region Responsible for Cadherin Internalization in Drosophila Melanogaster

• Main Author: 劉佑碩
• Other Authors: 徐瑞洲
• Format: Others
• Language: en_US
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/93569103238771765555

碩士 === 國立清華大學 === 分子醫學研究所 === 102 === E-cadherin is the major cell adhesion molecule (CAM) of adherens junctions (AJs), and it involves in many important cellular physiology. Echinoid (Ed), a nectin homologue, also belongs to CAM and localizes to AJs. To take a deeper look on the function of Ed in our previous study, we generated a chimeric construct which has an extracellular domain of Ed with an intracellular domain of cadherin (Ed-cad), and overexpressed Ed-cad in Drosophila tissue to see if it could replace the function of Ed or cadherin. Surprisingly, overexpression of Ed-cad led to endogenous cadherin endocytosis, so we looked for the factors which participate in cadherin internalization in this situation. Cadherin can be retrieved by different mechanism, mainly via clathrin-mediated endocytosis in most situation. However, there were still other unknown factors which involved in cadherin internalization when overexpressing Ed-cad. Overexpression of Ed-cad in adult Drosophila caused rough eye and wing defect. Here in this article, we used these features, and obtained Drosophila deficiency kit to generate a genome-wide screening to find out which factors may involve in cadherin internalization. After the screening, 7 genomic regions were identified, while some of them contain candidate genes which were already known or have the potential role involving in vesicular trafficking, including reck1, delta, sec10, Centaurin B1, lanB1, and fuzzy onion. Nonetheless, the study presented here is only the initial work to find out the unknown factors of cadherin recycling. In the future, we have to examine the candidate genes one by one by such as knock down assay, and need more biological evidence to prove the exactly roles of these specific genes.