Tuberin is a component of lipid rafts and mediates caveolin-1 localization: role of TSC2 in post-Golgi transport.
Mutations of the TSC2 gene lead to the development of hamartomas in tuberous sclerosis complex. Their pathology exhibits features indicative of defects in cell growth, proliferation, differentiation, and migration. We have previously shown that tuberin, the TSC2 protein, resides in multiple subcellular compartments and as such may serve multiple functions. To further characterize the microsomal pool of tuberin, we found that it cofractionated with caveolin-1 in a low-density, Triton X-100-resistant fraction (i.e., lipid rafts) and regulated its localization. In cells lacking tuberin, most of the endogenous caveolin-1 was displaced from the plasma membrane to a Brefeldin-A-sensitive, post-Golgi compartment distinct from the endosome and lysosome. Correspondingly, there was a paucity of caveolae at the plasma membrane of Tsc2-/- cells. Reintroduction of TSC2, but not a disease-causing mutant, reversed the caveolin-1 localization to the membrane. Exogenously expressed caveolin-1-GFP and vesicular stomatitis virus G protein, VSVG-GFP in the Tsc2-/- cells failed to be transported to the plasma membrane and were retained in distinct post-Golgi vesicles. Our data suggest a role of tuberin in regulating post-Golgi transport without apparent effects on protein sorting. The presence of mislocalized proteins in Tsc2-/- cells may contribute to the abnormal signaling and cellular phenotype of tuberous sclerosis.
Jones
Kathryn A
KA
Department of Surgery, University of Washington, Seattle, WA 98195, USA.
Jiang
Xiuyun
X
Yamamoto
Yuji
Y
Yeung
Raymond S
RS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D019556:COS Cells; D051242:Caveolin 1; D022461:Caveolins / Q000378:metabolism* / Q000648:ultrastructure; D002451:Cell Compartmentation; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003513:Cycloheximide / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000378:metabolism*; D021962:Membrane Microdomains / Q000737:chemistry*; D008861:Microsomes / Q000737:chemistry / Q000378:metabolism; D009154:Mutation; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D021381:Protein Transport; D011506:Proteins / Q000378:metabolism; D051381:Rats; D012097:Repressor Proteins / Q000378:metabolism*; D013696:Temperature; D000077004:Tuberous Sclerosis Complex 1 Protein; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics
D000911:Antibodies, Monoclonal; C495537:Cav1 protein, rat; D051242:Caveolin 1; D022461:Caveolins; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D012097:Repressor Proteins; C000624655:Tsc2 protein, rat; D000077004:Tuberous Sclerosis Complex 1 Protein; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins; D003513:Cycloheximide