title: BMCC1 is an AP-2 associated endosomal protein in prostate cancer cells.
abstract: The prostate cancer antigen gene 3 (PCA3) is embedded in an intron of a second gene BMCC1 (Bcl2-/adenovirus E1B nineteen kDa-interacting protein 2 (BNIP-2) and Cdc42GAP homology BCH motif-containing molecule at the carboxyl terminal region 1) which is also upregulated in prostate cancer. BMCC1 was initially annotated as two genes (C9orf65/PRUNE and BNIPXL) on either side of PCA3 but our data suggest that it represents a single gene coding for a high molecular weight protein. Here we demonstrate for the first time the expression of a >300 kDa BMCC1 protein (BMCC1-1) in prostate cancer and melanoma cell lines. This protein was found exclusively in the microsomal fraction and localised to cytoplasmic vesicles. We also observed expression of BMCC1 protein in prostate cancer sections using immunohistology. GST pull down, immunoprecipitation and mass spectrometry protein interaction studies identified multiple members of the Adaptor Related Complex 2 (AP-2) as BMCC1 interactors. Consistent with a role for BMCC1 as an AP-2 interacting endosomal protein, BMCC1 co-localised with β-adaptin at the perinuclear region of the cell. BMCC1 also showed partial co-localisation with the early endosome small GTP-ase Rab-5 as well as strong co-localisation with internalised pulse-chase labelled transferrin (Tf), providing evidence that BMCC1 is localised to functional endocytic vesicles. BMCC1 knockdown did not affect Tf uptake and AP-2 knockdown did not disperse BMCC1 vesicular distribution, excluding an essential role for BMCC1 in canonical AP-2 mediated endocytic uptake. Instead, we posit a novel role for BMCC1 in post-endocytic trafficking. This study provides fundamental characterisation of the BMCC1 complex in prostate cancer cells and for the first time implicates it in vesicle trafficking.
authors.affiliation: Queensland Institute of Medical Research, Herston, Brisbane, Queensland, Australia.
authors.forename: Janelle L
authors.initials: JL
authors.lastname: Harris
authors.forename: Renée S
authors.initials: RS
authors.lastname: Richards
authors.forename: Clement W K
authors.initials: CW
authors.lastname: Chow
authors.forename: Soon
authors.initials: S
authors.lastname: Lee
authors.forename: Misook
authors.initials: M
authors.lastname: Kim
authors.forename: Marion
authors.initials: M
authors.lastname: Buck
authors.forename: Linda
authors.initials: L
authors.lastname: Teng
authors.forename: Raymond
authors.initials: R
authors.lastname: Clarke
authors.forename: Robert A
authors.initials: RA
authors.lastname: Gardiner
authors.forename: Martin F
authors.initials: MF
authors.lastname: Lavin
mesh_terms: D033962:Adaptor Protein Complex 2 / Q000378:metabolism*; D000818:Animals; D045744:Cell Line, Tumor; D011992:Endosomes / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D008297:Male; D051379:Mice; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D009928:Organ Specificity / Q000235:genetics; D011471:Prostatic Neoplasms / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D021381:Protein Transport; D022161:Transport Vesicles / Q000378:metabolism
chemical_list: D033962:Adaptor Protein Complex 2; D009363:Neoplasm Proteins; C509845:PRUNE2 protein, human
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