The AE2 anion exchanger is necessary for the structural integrity of the Golgi apparatus in mammalian cells.
The structural integrity of the Golgi apparatus is known to be dependent on multiple factors, including the organizational status of microtubules, actin and the ankyrin/spectrin-based Golgi membrane skeleton, as well as vesicular trafficking and pH homeostasis. In this respect, our recently identified Golgi-associated anion exchanger, AE2, may also be of importance, since it potentially acts as a Golgi pH regulator and as a novel membrane anchor for the spectrin-based Golgi membrane skeleton. Here, we show that inhibition (>75%) of AE2 expression by antisense oligonucleotides in COS-7 cells results in the fragmentation of the juxtanuclear Golgi apparatus and in structural disorganization of the Golgi stacks, the cisternae becoming generally shorter, distorted, vesiculated and/or swollen. These structural changes occurred without apparent dissociation of the Golgi membrane skeletal protein Ankyrin(195), but were accompanied by the disappearance of the well-focused microtubule-organizing center (MTOC), suggesting the involvement of microtubule reorganization. Similar changes in Golgi structure and assembly of the MTOC were also observed upon transient overexpression of the EGFP-AE2 fusion protein. These data implicate a clear structural role for the AE2 protein in the Golgi and in its cytological positioning around the MTOC.
Holappa
Katja
K
Department of Biochemistry, University of Oulu, P.O. Box 3000, 90014 Oulu, Finland.
Muñoz
Maria Teresa
MT
Egea
Gustavo
G
Kellokumpu
Sakari
S
D000818:Animals; D027321:Anion Transport Proteins; D017487:Ankyrins; D017920:Antiporters; D019556:COS Cells; D002522:Chlorocebus aethiops; D006056:Golgi Apparatus / Q000648:ultrastructure*; D007425:Intracellular Membranes / Q000737:chemistry / Q000648:ultrastructure; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D022101:Microtubule-Organizing Center / Q000648:ultrastructure; D008870:Microtubules / Q000648:ultrastructure; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D064606:SLC4A Proteins; D014162:Transfection
D027321:Anion Transport Proteins; D017487:Ankyrins; D017920:Antiporters; D008565:Membrane Proteins; D016376:Oligonucleotides, Antisense; D064606:SLC4A Proteins