title: Golgb1 regulates protein glycosylation and is crucial for mammalian palate development.
abstract: Cleft palate is a common major birth defect for which currently known causes account for less than 30% of pathology in humans. In this study, we carried out mutagenesis screening in mice to identify new regulators of palatogenesis. Through genetic linkage mapping and whole-exome sequencing, we identified a loss-of-function mutation in the Golgb1 gene that co-segregated with cleft palate in a new mutant mouse line. Golgb1 is a ubiquitously expressed large coiled-coil protein, also known as giantin, that is localized at the Golgi membrane. Using CRISPR/Cas9-mediated genome editing, we generated and analyzed developmental defects in mice carrying additional Golgb1 loss-of-function mutations, which supported a crucial requirement for Golgb1 in palate development. Through maxillary explant culture assays, we demonstrate that the Golgb1 mutant embryos have intrinsic defects in palatal shelf elevation. Just prior to the developmental stage of palatal shelf elevation in wild-type littermates, Golgb1 mutant embryos exhibit increased cell density, reduced hyaluronan accumulation and impaired protein glycosylation in the palatal mesenchyme. Together, these results demonstrate that, although it is a ubiquitously expressed Golgi-associated protein, Golgb1 has specific functions in protein glycosylation and tissue morphogenesis.
authors.affiliation: Division of Plastic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA Yu.Lan@cchmc.org.
authors.forename: Yu
authors.identifier: 0000-0001-5611-3600
authors.initials: Y
authors.lastname: Lan
authors.affiliation: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Nian
authors.initials: N
authors.lastname: Zhang
authors.affiliation: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Han
authors.initials: H
authors.lastname: Liu
authors.affiliation: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Jingyue
authors.initials: J
authors.lastname: Xu
authors.affiliation: Division of Plastic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Rulang
authors.initials: R
authors.lastname: Jiang
mesh_terms: D000818:Animals; D001483:Base Sequence; D064113:CRISPR-Cas Systems / Q000235:genetics; D002452:Cell Count; D002972:Cleft Palate / Q000235:genetics; D004622:Embryo, Mammalian / Q000378:metabolism; D005038:Ethylnitrosourea; D005260:Female; D005455:Fluorescent Antibody Technique; D016678:Genome; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism / Q000648:ultrastructure; D000077614:Golgi Matrix Proteins; D006820:Hyaluronic Acid / Q000378:metabolism; D037102:Lectins / Q000378:metabolism; D008322:Mammals / Q000196:embryology*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008648:Mesoderm / Q000166:cytology / Q000196:embryology; D008817:Mice, Mutant Strains; D009154:Mutation / Q000235:genetics; D010159:Palate / Q000196:embryology* / Q000378:metabolism*; D017393:RNA Editing / Q000235:genetics; D022821:RNA Splice Sites / Q000235:genetics; D012326:RNA Splicing / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D000077614:Golgi Matrix Proteins; D037102:Lectins; D008565:Membrane Proteins; D022821:RNA Splice Sites; D012333:RNA, Messenger; C087574:macrogolgin; D006820:Hyaluronic Acid; D005038:Ethylnitrosourea
keywords: CRISPR; Cleft palate; Craniofacial; ENU; Genome editing; Glycosylation; Golgi; Mutation