title: Absence of post-translational aspartyl beta-hydroxylation of epidermal growth factor domains in mice leads to developmental defects and an increased incidence of intestinal neoplasia.
abstract: The BAH genomic locus encodes three distinct proteins: junctin, humbug, and BAH. All three proteins share common exons, but differ significantly based upon the use of alternative terminal exons. The biological roles of BAH and humbug and their functional relationship to junctin remain unclear. To evaluate the role of BAH in vivo, the catalytic domain of BAH was specifically targeted such that the coding regions of junctin and humbug remained undisturbed. BAH null mice lack measurable BAH protein in several tissues, lack aspartyl beta-hydroxylase activity in liver preparations, and exhibit no hydroxylation of the epidermal growth factor (EGF) domain of clotting Factor X. In addition to reduced fertility in females, BAH null mice display several developmental defects including syndactyly, facial dysmorphology, and a mild defect in hard palate formation. The developmental defects present in BAH null mice are similar to defects observed in knock-outs and hypomorphs of the Notch ligand Serrate-2. In this work, beta-hydroxylation of Asp residues in EGF domains is demonstrated for a soluble form of a Notch ligand, human Jagged-1. These results along with recent reports that another post-translational modification of EGF domains in Notch gene family members (glycosylation by Fringe) alters Notch pathway signaling, lends credence to the suggestion that aspartyl beta-hydroxylation may represent another post-translational modification of EGF domains that can modulate Notch pathway signaling. Previous work has demonstrated increased levels of BAH in certain tumor tissues and a role for BAH in tumorigenesis has been proposed. The role of hydroxylase in tumor formation was tested directly by crossing BAH KO mice with an intestinal tumor model, APCmin mice. Surprisingly, BAH null/APCmin mice show a statistically significant increase in both intestinal polyp size and number when compared with BAH wild-type/APCmin controls. These results suggest that, in contrast to expectations, loss of BAH catalytic activity may promote tumor formation.
authors.affiliation: Bristol-Myers Squibb Pharma Research Labs, Inc., Wilmington, DE 19880-0400, USA. joseph.dinchuk@bms.com
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Dinchuk
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Focht
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Kelley
authors.forename: Nancy L
authors.initials: NL
authors.lastname: Henderson
authors.forename: Nina I
authors.initials: NI
authors.lastname: Zolotarjova
authors.forename: Richard
authors.initials: R
authors.lastname: Wynn
authors.forename: Nicola T
authors.initials: NT
authors.lastname: Neff
authors.forename: John
authors.initials: J
authors.lastname: Link
authors.forename: Reid M
authors.initials: RM
authors.lastname: Huber
authors.forename: Timothy C
authors.initials: TC
authors.lastname: Burn
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Rupar
authors.forename: Mark R
authors.initials: MR
authors.lastname: Cunningham
authors.forename: Bernard H
authors.initials: BH
authors.lastname: Selling
authors.forename: Jianhong
authors.initials: J
authors.lastname: Ma
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Stern
authors.forename: Gregory F
authors.initials: GF
authors.lastname: Hollis
authors.forename: Robert B
authors.initials: RB
authors.lastname: Stein
authors.forename: Paul A
authors.initials: PA
authors.lastname: Friedman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D004815:Epidermal Growth Factor / Q000378:metabolism*; D005091:Exons; D005260:Female; D006900:Hydroxylation; D015994:Incidence; D007414:Intestinal Neoplasms / Q000235:genetics*; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D006899:Mixed Function Oxygenases / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D051880:Receptors, Notch
chemical_list: D008565:Membrane Proteins; D051880:Receptors, Notch; D004815:Epidermal Growth Factor; D006899:Mixed Function Oxygenases; C058291:aspartic acid 2-oxoglutarate-dependent dioxygenase