A critical role of cavin (polymerase I and transcript release factor) in caveolae formation and organization.
Cavin (PTRF) has been shown to be a highly abundant protein component of caveolae, but its functional role there is unknown. Here, we confirm that cavin co-localizes with caveolin-1 in adipocytes by confocal microscopy and co-distributes with caveolin-1 in lipid raft fractions by sucrose gradient flotation. However, cavin does not directly associate with caveolin-1 as solubilization of caveolae disrupts their interaction. Cholesterol depletion with beta-cyclodextrin causes a significant down-regulation of cavin from plasma membrane lipid raft fractions. Overexpression of cavin in HEK293-Cav-1 cells and knockdown of cavin in 3T3-L1 adipocytes enhances and diminishes caveolin-1 levels, respectively, indicating an important role for cavin in maintaining the level of caveolin-1. A truncated form of cavin, eGFP-cavin-1-322, which lacks 74 amino acids from the C-terminal, reveals a microtubular network localization by confocal microscopy. Disruption of cytoskeletal elements with latrunculin B or nocodazole diminishes cavin expression without affecting the caveolin-1 amount. We propose that the presence of cavin on the inside surface of caveolae stabilizes these structures, probably through interaction with the cytoskeleton, and cavin therefore plays an important role in caveolae formation and organization.
Liu
Libin
L
Department of Biochemistry, Boston University Medical School, Boston, Massachusetts 02118, USA.
Pilch
Paul F
PF
D041721:3T3-L1 Cells; D000199:Actins / Q000378:metabolism; D017667:Adipocytes / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D021941:Caveolae; D002460:Cell Line; D017931:DNA Primers; D006801:Humans; D047468:Immunoprecipitation; D008565:Membrane Proteins / Q000502:physiology*; D051379:Mice; D018613:Microscopy, Confocal; D008870:Microtubules / Q000378:metabolism; D034622:RNA Interference; D016601:RNA-Binding Proteins
D000199:Actins; C112745:Cavin1 protein, mouse; D017931:DNA Primers; D008565:Membrane Proteins; D016601:RNA-Binding Proteins