MCDB428Lect14 - MCDB 428 Lecture 14: Friday February 8,...

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Lecture 14: Friday February 8, 2008 Today’s Outline Brefeldin A Effect on membrane trafficking Mechanism of action Clathrin Coated Vesicles Different types of CCVs Roles in post-Golgi trafficking Formation of CCVs Role of Dynamin Shibire mutant Drosphila Effect of dynamin mutations on CCV formation Retromer Coats Lysosomal sorting receptor pathway Retrieval of sorting receptors Formation of the retromer coat Involvement of PI-3P binding domain The phosphoinositide Family Description of PI and PI-isoforms PI-isoform family complement How PI-binding domains display specificity Where different PI-isoforms are distributed intracellularly MCDB 428
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Brefeldin A Causes Loss of Organelle Identity Brefeldin A (BFA) is a lactone antibiotic produced by certain fungi Treatment of eukaryotic cells with BFA results in rapid inhibition of membrane trafficking This membrane trafficking inhibition dramatically causes dissolution of the Golgi complex Golgi breakdown initiates with formation of extended tubules from Golgi cisternae This is rapidly followed by accumulation of Golgi resident proteins in the ER GFP-galactosyltransferase – a trans-Golgi resident enzyme
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BFA Interferes with ArfGEF Activities BFA binds to ArfGEFs containing a Sec7 homology domain This binding uncompetitively inhibits Arf1 GTPase binding to the ArfGEF As a result Arf1GTPases are unable to become activated on the Golgi and as a result intra Golgi transport is inhibited as well as retrograde trafficking to the ER This blockage of trafficking results in loss of organelle identity and eventually breakdown of trafficking between the ER and Golgi
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Clathrin Coated Vesicles (CCVs) Clathrin coated vesicles (CCVs) involved in various post-Golgi trafficking steps Clathrin is recruited by different types of adaptor proteins, AP-1 (Golgi-Endosome), AP-2 (Plasma Membrane-Endosome), GGA (Golgi-Endosome), AP-3 (Golgi-Lysosome)
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Different Adaptor Complexes Target Clathrin Coated Vesicles to Different Compartments
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This note was uploaded on 09/03/2009 for the course MCDB 428 taught by Professor Wang during the Winter '08 term at University of Michigan.

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MCDB428Lect14 - MCDB 428 Lecture 14: Friday February 8,...

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