ZFIN ID: ZDB-FIG-160511-11
DeRossi et al., 2016 - trappc11 is required for protein glycosylation in zebrafish and humans. Molecular biology of the cell   27(8):1220-34 Full text @ Mol. Biol. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Terms:
Stage: Day 5
PHENOTYPE:
Fish:
Observed In:
Stage: Day 5

Fig. 5

Terpenoid and dolichol synthesis defects in trappc11 mutants. (A) Schematic representation of the terpenoid biosynthetic pathway, with branch points at farnesyl-PP leading to production of dolichol (protein glycosylation), squalene (steroid synthesis), and geranylgeranyl-PP (protein prenylation). Fold changes of expression in trappc11 mutant livers are indicated by color: blue indicates decrease and red indicates increase relative to WT siblings; gray indicates genes not tested. Drugs used are indicated with a red T at their targets. (B) Confocal microscopy of transgenic WT and trappc11-mutant zebrafish expressing a prenylated form of GFP (Tg(actb2:CAAX-GFP)) in hepatocytes. Zebrafish were collected at 5 dpf and analyzed for membrane localization of CAAX-GFP. Samples were counterstained with DAPI (nucleus) and Cy5-streptavidin to stain the biotin-rich hepatocytes (middle; white boxes outline magnified GFP-only images shown on right). Scale bars, 10 µm. (C) Confocal microscopy of WT transgenic zebrafish expressing a prenylated form of GFP in hepatocytes (Tg(fabp10:CAAX-GFP). Zebrafish larvae were treated with DMSO (top) or 0.75 µg/ml BFA (bottom) from 3 to 5 dpf and analyzed at 5 dpf for membrane localization of CAAX-GFP. Samples were counterstained with DAPI (blue nucleus). White boxes outline area shown magnified on right (GFP only). Scale bars, 10 µM. (D) Whole WT and trappc11-mutant larvae were collected at 5 dpf, and total cholesterol was measured and normalized to protein content. trappc11 mutants have a significant decrease in cholesterol compared with WT siblings (p = 0.0008, paired Student’s t test), suggestive of decreased terpenoid biosynthetic activity. Individual clutches are indicated (n = 17 for each), with lines representing clutch pairing.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
alg2 trappc11hi1532bTg/hi1532bTg standard conditions Day 5 liver RTPCR
dhdds trappc11hi1532bTg/hi1532bTg standard conditions Day 5 liver RTPCR
dpagt1 trappc11hi1532bTg/hi1532bTg standard conditions Day 5 liver RTPCR
dpm1 trappc11hi1532bTg/hi1532bTg standard conditions Day 5 liver RTPCR
dpm2 trappc11hi1532bTg/hi1532bTg standard conditions Day 5 liver RTPCR
dpm3 trappc11hi1532bTg/hi1532bTg standard conditions Day 5 liver RTPCR
fdft1 trappc11hi1532bTg/hi1532bTg standard conditions Day 5 liver RTPCR
GFP mss11Tg standard conditions Day 5 hepatocyte plasma membrane IFL
mss11Tg chemical treatment: brefeldin A Day 5 hepatocyte plasma membrane IFL
trappc11hi1532bTg/hi1532bTg; mss11Tg standard conditions Day 5 hepatocyte cytoplasm IFL
Day 5 hepatocyte plasma membrane IFL
gmppb trappc11hi1532bTg/hi1532bTg standard conditions Day 5 liver RTPCR
mvda trappc11hi1532bTg/hi1532bTg standard conditions Day 5 liver RTPCR
pmm2 trappc11hi1532bTg/hi1532bTg standard conditions Day 5 liver RTPCR
srd5a3 trappc11hi1532bTg/hi1532bTg standard conditions Day 5 liver RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
trappc11hi1532bTg/hi1532bTg standard conditions Day 5 liver alg2 expression increased amount, abnormal
Day 5 liver dhdds expression increased amount, abnormal
Day 5 liver dpagt1 expression increased amount, abnormal
Day 5 liver dpm1 expression increased amount, abnormal
Day 5 liver dpm2 expression increased amount, abnormal
Day 5 liver dpm3 expression increased amount, abnormal
Day 5 liver fdft1 expression decreased amount, abnormal
Day 5 liver gmppb expression increased amount, abnormal
Day 5 liver mvda expression increased amount, abnormal
Day 5 liver pmm2 expression increased amount, abnormal
Day 5 liver srd5a3 expression increased amount, abnormal
trappc11hi1532bTg/hi1532bTg; mss11Tg standard conditions Day 5 cholesterol homeostasis disrupted, abnormal
Day 5 protein secretion disrupted, abnormal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Mol. Biol. Cell