title: The endocytic adaptor protein ARH associates with motor and centrosomal proteins and is involved in centrosome assembly and cytokinesis.
abstract: Numerous proteins involved in endocytosis at the plasma membrane have been shown to be present at novel intracellular locations and to have previously unrecognized functions. ARH (autosomal recessive hypercholesterolemia) is an endocytic clathrin-associated adaptor protein that sorts members of the LDL receptor superfamily (LDLR, megalin, LRP). We report here that ARH also associates with centrosomes in several cell types. ARH interacts with centrosomal (gamma-tubulin and GPC2 and GPC3) and motor (dynein heavy and intermediate chains) proteins. ARH cofractionates with gamma-tubulin on isolated centrosomes, and gamma-tubulin and ARH interact on isolated membrane vesicles. During mitosis, ARH sequentially localizes to the nuclear membrane, kinetochores, spindle poles and the midbody. Arh(-/-) embryonic fibroblasts (MEFs) show smaller or absent centrosomes suggesting ARH plays a role in centrosome assembly. Rat-1 fibroblasts depleted of ARH by siRNA and Arh(-/-) MEFs exhibit a slower rate of growth and prolonged cytokinesis. Taken together the data suggest that the defects in centrosome assembly in ARH depleted cells may give rise to cell cycle and mitotic/cytokinesis defects. We propose that ARH participates in centrosomal and mitotic dynamics by interacting with centrosomal proteins. Whether the centrosomal and mitotic functions of ARH are related to its endocytic role remains to be established.
authors.affiliation: Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA 92093, USA.
authors.forename: Sanna
authors.initials: S
authors.lastname: Lehtonen
authors.forename: Mehul
authors.initials: M
authors.lastname: Shah
authors.forename: Rikke
authors.initials: R
authors.lastname: Nielsen
authors.forename: Noriaki
authors.initials: N
authors.lastname: Iino
authors.forename: Jennifer J
authors.initials: JJ
authors.lastname: Ryan
authors.forename: Huilin
authors.initials: H
authors.lastname: Zhou
authors.forename: Marilyn G
authors.initials: MG
authors.lastname: Farquhar
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D018385:Centrosome / Q000378:metabolism*; D048749:Cytokinesis; D004705:Endocytosis; D005347:Fibroblasts / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000378:metabolism; D051379:Mice; D011485:Protein Binding; D051381:Rats; D014404:Tubulin / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C469745:LDLRAP1 protein, human; C500241:Ldlrap1 protein, mouse; D014404:Tubulin