PUBLICATION
The C-mannosylome of human induced pluripotent stem cells implies a role for ADAMTS16 C-mannosylation in eye development
- Authors
- Cirksena, K., Hütte, H.J., Shcherbakova, A., Thumberger, T., Sakson, R., Weiss, S., Jensen, L.R., Friedrich, A., Todt, D., Kuss, A.W., Ruppert, T., Wittbrodt, J., Bakker, H., Buettner, F.F.R.
- ID
- ZDB-PUB-210512-15
- Date
- 2021
- Source
- Molecular & cellular proteomics : MCP 20: 100092 (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
-
- Mannose
- Mannosyltransferases/genetics
- Mannosyltransferases/metabolism*
- Eye/growth & development*
- Humans
- Gene Editing
- Cell Line
- Induced Pluripotent Stem Cells/metabolism
- Oryzias
- ADAMTS Proteins/metabolism*
- Gene Knockdown Techniques
- Animals
- Cricetulus
- PubMed
- 33975020 Full text @ Mol. Cell. Proteomics
Citation
Cirksena, K., Hütte, H.J., Shcherbakova, A., Thumberger, T., Sakson, R., Weiss, S., Jensen, L.R., Friedrich, A., Todt, D., Kuss, A.W., Ruppert, T., Wittbrodt, J., Bakker, H., Buettner, F.F.R. (2021) The C-mannosylome of human induced pluripotent stem cells implies a role for ADAMTS16 C-mannosylation in eye development. Molecular & cellular proteomics : MCP. 20:100092.
Abstract
C-mannosylation is a modification of tryptophan residues with a single mannose, and can affect protein folding, secretion and/or function. To date, only a few proteins have been demonstrated to be C-mannosylated, and studies that globally assess protein C-mannosylation are scarce. To interrogate the C-mannosylome of human induced pluripotent stem cells (hiPSCs), we compared the secretomes of CRISPR-Cas9 mutants lacking either the C-mannosyltransferase DPY19L1 or DPY19L3 to wild-type hiPSCs using mass spectrometry-based quantitative proteomics. The secretion of numerous proteins was reduced in these mutants, including that of ADAMTS16, an extracellular protease that was previously reported to be essential for optic fissure fusion in zebrafish eye development. To test the functional relevance of this observation, we targeted dpy19l1 or dpy19l3 in embryos of the Japanese rice fish medaka (Oryzias latipes) by CRISPR-Cas9. We observed that targeting of dpy19l3 partially caused defects in optic fissure fusion, called coloboma. We further showed in a cellular model that DPY19L1 and DPY19L3 mediate C-mannosylation of a recombinantly expressed thrombospondin type 1 repeat of ADAMTS16 and thereby support its secretion. Taken together, our findings imply that DPY19L3-mediated C-mannosylation is involved in eye development by assisting secretion of the extracellular protease ADAMTS16.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping