李芳仁2006-07-262018-07-092006-07-262018-07-092003-07-31http://ntur.lib.ntu.edu.tw//handle/246246/23778To identify molecules that might act as down-stream effectors of hARL1, we used hARL1(Q/L) as bait in a yeast two-hybrid screen of human fetal liver cDNA library. 43 positives were obtained and several distinct genes were chosen for further analysis. Among them, golgin-245 (9 clones), ApoC2 (8 clones), Arfaptin 2 / POR1 (4 clones), ApoB (2 clones), and Sec10 (one clone) will be further characterized to support the their functional interactions. We also constructed wild type hARL1 and hARL1-T/N to test whether interaction of hARL1 and their interacting proteins is nucleotide-dependent. We also found that hARL1 and golgin-245 bound to Golgi membranes, consistent with a function in vesicular trafficking. Expression of putatively constitutively active mutant of hARL1, ARL1(Q71L), in cells led to down-regulate golgin-245 bound to Golgi membranes. In vitro protein-interaction assays showed that hARL1(Q71L) interacted with C-terminal of golgin-245. How is sorting signal recognition regulated so that interaction with each vesicle-receptor binding protein occurs only at the appropriate organelle? Cooperative association of cargo, vesicle binding proteins, and additional factors into mutimeric, trans-Golgi and/or lysosome-specific complexes will be investigated in this project.application/pdf158470 bytesapplication/pdfzh-TW國立臺灣大學醫學院分子醫學研究所journal articlehttp://ntur.lib.ntu.edu.tw/bitstream/246246/23778/1/913112B002023.pdf