ADRP is dissociated from lipid droplets by ARF1-dependent mechanism.
Adipocyte differentiation-related protein (ADRP) is a member of PAT proteins existing in lipid droplets (LDs). By yeast two-hybrid screening, we identified ADP-ribosylation factor 1 (ARF1) as a binding partner of ADRP. The interaction of ADRP and ARF1 was verified by GST pull-down and co-immunoprecipitation experiments. Interestingly, ADRP precipitated the GDP-bound ARF1 preferentially to the GTP-bound ARF1. Consistent with this, either brefeldin A (BFA), a fungal metabolite to inhibit ARF-GEF, or a dominant-negative mutant of ARF1 caused dissociation of ADRP from LD. On the other hand, overexpression of wild-type ARF1 did not promote the ADRP dissociation or new LD formation. By using deletion mutants, a central domain of ADRP, which is dispensable for LD binding, was shown to bind to ARF1. The present study showed that the GDP-bound ARF1 induces dissociation of ADRP from the LD surface, and that LD is a target of BFA action.
Nakamura
Noriko
N
Department of Anatomy and Molecular Cell Biology, Nagoya University, Graduate School of Medicine, 65 Tsurumai, Showa, Nagoya 466-8550, Japan. noriko@med.nagoya-u.ac.jp
Akashi
Tomohiro
T
Taneda
Tsuya
T
Kogo
Hiroshi
H
Kikuchi
Akihiko
A
Fujimoto
Toyoshi
T
D016475:3T3 Cells; D020823:ADP-Ribosylation Factor 1 / Q000502:physiology*; D000818:Animals; D020126:Brefeldin A / Q000494:pharmacology; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D006153:Guanosine Diphosphate / Q000378:metabolism; D006801:Humans; D008055:Lipids / Q000502:physiology; D008565:Membrane Proteins / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D000070780:Perilipin-2; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism
D008055:Lipids; D008565:Membrane Proteins; C000602982:PLIN2 protein, human; D000070780:Perilipin-2; C000602983:Plin2 protein, mouse; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D006153:Guanosine Diphosphate; D020126:Brefeldin A; D020823:ADP-Ribosylation Factor 1