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Functional characterization of zebrafish orthologs of the human Beta 3-Glucosyltransferase B3GLCT gene mutated in Peters Plus Syndrome
- Source :
- PLoS ONE, Vol 12, Iss 9, p e0184903 (2017), PLoS ONE
- Publication Year :
- 2017
- Publisher :
- Public Library of Science (PLoS), 2017.
-
Abstract
- Peters Plus Syndrome (PPS) is a rare autosomal recessive disease characterized by ocular defects, short stature, brachydactyly, characteristic facial features, developmental delay and other highly variable systemic defects. Classic PPS is caused by loss-of-function mutations in the B3GLCT gene encoding for a β3-glucosyltransferase that catalyzes the attachment of glucose via a β1–3 glycosidic linkage to O-linked fucose on thrombospondin type 1 repeats (TSRs). B3GLCT was shown to participate in a non-canonical ER quality control mechanism; however, the exact molecular processes affected in PPS are not well understood. Here we report the identification and characterization of two zebrafish orthologs of the human B3GLCT gene, b3glcta and b3glctb. The b3glcta and b3glctb genes encode for 496-aa and 493-aa proteins with 65% and 57% identity to human B3GLCT, respectively. Expression studies demonstrate that both orthologs are widely expressed with strong presence in embryonic tissues affected in PPS. In vitro glucosylation assays demonstrated that extracts from wildtype embryos contain active b3glct enzyme capable of transferring glucose from UDP-glucose to an O-fucosylated TSR, indicating functional conservation with human B3GLCT. To determine the developmental role of the zebrafish genes, single and double b3glct knockouts were generated using TALEN-induced genome editing. Extracts from double homozygous b3glct-/- embryos demonstrated complete loss of in vitro b3glct activity. Surprisingly, b3glct-/- homozygous fish developed normally. Transcriptome analyses of head and trunk tissues of b3glct-/- 24-hpf embryos identified 483 shared differentially regulated transcripts that may be involved in compensation for b3glct function in these embryos. The presented data show that both sequence and function of B3GLCT/b3glct genes is conserved in vertebrates. At the same time, complete b3glct deficiency in zebrafish appears to be inconsequential and possibly compensated for by a yet unknown mechanism.
- Subjects :
- Genome engineering
0301 basic medicine
Embryology
Embryo, Nonmammalian
lcsh:Medicine
Engineering and technology
Synthetic genome editing
medicine.disease_cause
Cornea
Transcriptome
Gene Knockout Techniques
Database and Informatics Methods
0302 clinical medicine
Synthetic bioengineering
lcsh:Science
Zebrafish
Growth Disorders
In Situ Hybridization
Gene Editing
Genetics
Mutation
Mammalian Genomics
Multidisciplinary
Eukaryota
Animal Models
Genomics
TALENs
Experimental Organism Systems
Glucosyltransferases
Osteichthyes
Vertebrates
Sequence Analysis
Research Article
Biotechnology
Quality Control
Bioinformatics
Cleft Lip
Molecular Sequence Data
Limb Deformities, Congenital
Bioengineering
Biology
Research and Analysis Methods
03 medical and health sciences
Model Organisms
Sequence Motif Analysis
Transcription Activator-Like Effector Nucleases
Industrial Engineering
medicine
Animals
Humans
Adults
Amino Acid Sequence
Gene
Synthetic biology
Gene knockout
Gene Expression Profiling
Embryos
Synthetic genomics
lcsh:R
Organisms
Wild type
Biology and Life Sciences
Zebrafish Proteins
biology.organism_classification
Gene expression profiling
Fish
030104 developmental biology
Animal Genomics
Age Groups
People and Places
Population Groupings
lcsh:Q
Sequence Alignment
030217 neurology & neurosurgery
Developmental Biology
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 12
- Issue :
- 9
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....92178665ab1117d50e77d5a7137f78c5