title: Post-translational regulation of CD133 by ATase1/ATase2-mediated lysine acetylation.
abstract: The CD133 cell-surface protein expresses the AC133 epitope that is associated with cancer progenitor cells and cancer resistance to traditional anticancer therapies. We report that the endoplasmic reticulum Golgi intermediate compartment residing acetyltransferases, ATase1 (NAT8B) and ATase2 (NAT8), can physically interact with CD133 to acetylate the protein on three lysine residues predicted to reside on the first extracellular loop of CD133. Site-directed mutagenesis of these residues mimicking a loss of acetylation and downregulation or inhibition of ATase1/ATase2 resulted in near-complete abolishment of CD133 protein expression. We also demonstrate that targeting ATase1/ATase2 results in apoptosis of CD133 expressing acute lymphoblastic leukemia cells. Taken together, we suggest that lysine acetylation on predicted extracellular residues plays a key role in expression and trafficking of CD133 protein to the cell surface and can be targeted to disrupt CD133 regulation and function.
authors.affiliation: Donnelly Centre and Banting and Best Department of Medical Research, University of Toronto, 160 College Street, Toronto, ON M5S 1A8, Canada; Department of Molecular Genetics, University of Toronto, TorontoON M5S 1A8, Canada.
authors.forename: Anthony B
authors.initials: AB
authors.lastname: Mak
authors.affiliation: Department of Medicine, University of Wisconsin-Madison, 2500 Overlook Terrace, Madison, WI 53705, USA.
authors.forename: Mariana
authors.initials: M
authors.lastname: Pehar
authors.affiliation: Donnelly Centre and Banting and Best Department of Medical Research, University of Toronto, 160 College Street, Toronto, ON M5S 1A8, Canada; Department of Molecular Genetics, University of Toronto, TorontoON M5S 1A8, Canada.
authors.forename: Allison M L
authors.initials: AM
authors.lastname: Nixon
authors.affiliation: Donnelly Centre and Banting and Best Department of Medical Research, University of Toronto, 160 College Street, Toronto, ON M5S 1A8, Canada; Department of Molecular Genetics, University of Toronto, TorontoON M5S 1A8, Canada.
authors.forename: Rashida A
authors.initials: RA
authors.lastname: Williams
authors.affiliation: Donnelly Centre and Banting and Best Department of Medical Research, University of Toronto, 160 College Street, Toronto, ON M5S 1A8, Canada; Department of Molecular Genetics, University of Toronto, TorontoON M5S 1A8, Canada.
authors.forename: Andrea C
authors.initials: AC
authors.lastname: Uetrecht
authors.affiliation: Department of Medicine, University of Wisconsin-Madison, 2500 Overlook Terrace, Madison, WI 53705, USA; Geriatric Research Education Clinical Center, VA Medical Center, 2500 Overlook Terrace, Madison, WI 53705, USA.
authors.forename: Luigi
authors.initials: L
authors.lastname: Puglielli
authors.affiliation: Donnelly Centre and Banting and Best Department of Medical Research, University of Toronto, 160 College Street, Toronto, ON M5S 1A8, Canada; Department of Molecular Genetics, University of Toronto, TorontoON M5S 1A8, Canada. Electronic address: j.moffat@utoronto.ca.
authors.forename: Jason
authors.initials: J
authors.lastname: Moffat
mesh_terms: D000071916:AC133 Antigen; D000107:Acetylation / Q000187:drug effects; D000123:Acetyltransferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D018938:Caco-2 Cells; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D008239:Lysine / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D014407:Tumor Cells, Cultured
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D004791:Enzyme Inhibitors; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D000123:Acetyltransferases; C116373:NAT8 protein, human; D008239:Lysine
keywords: ERGIC; membrane protein; post-translation modifications; protein processing; trafficking