Projects per year
Abstract
Post-translational modification of serine/threonine residues in nucleocytoplasmic proteins with N-acetylglucosamine (OGlcNAcylation) is an essential regulatory mechanism in many cellular processes. In Drosophila, null mutants of the polycomb gene O-GlcNAc transferase (OGT, also known as super sex combs [sxc]) display homeotic phenotypes. To dissect the requirement for OGlcNAc signaling in Drosophila development, we used CRISPR/Cas9 gene editing to generate rationally designed sxc catalytically hypomorphic or null point mutants. Of the fertile males derived from embryos injected with the CRISPR/Cas9 reagents, 25% produced progeny carrying precise point mutations with no detectable off-target effects. One of these mutants, the catalytically inactive sxcK872M, was recessive lethal, whereas a second mutant, the hypomorphic sxcH537A, was homozygous viable. We observed that reduced total protein O-GlcNAcylation in the sxcH537A mutant is associated with a wing vein phenotype and temperature-dependent lethality. Genetic interaction between sxcH537A and a null allele of Drosophila host cell factor (dHcf), encoding an extensively O-GlcNAcylated transcriptional coactivator, resulted in abnormal scutellar bristle numbers. A similar phenotype was also observed in sxcH537A flies lacking a copy of skuld (skd), a Mediator complex gene known to affect scutellar bristle formation. Interestingly, this phenotype was independent of OGT Polycomb function or dHcf downstream targets. In conclusion, the generation of the endogenous OGT hypomorphic mutant sxcH537A enabled us to identify pleiotropic effects of globally reduced protein O-GlcNAc during Drosophila development. The mutants generated and phenotypes observed in this study provide a platform for discovery of OGT substrates that are critical for Drosophila development.
Original language | English |
---|---|
Pages (from-to) | 7209-7221 |
Number of pages | 13 |
Journal | Journal of Biological Chemistry |
Volume | 293 |
Issue number | 19 |
Early online date | 27 Mar 2018 |
DOIs | |
Publication status | Published - 11 May 2018 |
Keywords
- O-GlcNAc
- O-GlcNAc transferase
- host cell factor
- CRISPR/Cas9
- Drosophila development
- polycomb
- Mediator complex
- post-translational protein modification
- hypomorphic sxc mutant
ASJC Scopus subject areas
- Molecular Biology
- Biochemistry
- Cell Biology
Fingerprint
Dive into the research topics of 'Effects of hypo O-GlcNAcylation on Drosophila development'. Together they form a unique fingerprint.Projects
- 2 Finished
-
Molecular Mechanisms of O-GICNAC Signalling (Investigator award)
van Aalten, D. (Investigator)
1/03/16 → 28/02/22
Project: Research
-
Dynamics of Fundamental Cellular Processes by Live Cell and Tissue Imaging
MacDonald, M. (Investigator), McGloin, D. (Investigator), McKenna, S. (Investigator), Storey, K. (Investigator), Swedlow, J. (Investigator) & Weijer, K. (Investigator)
1/01/13 → 31/12/17
Project: Research
Profiles
-
van Aalten, Daan
- Molecular Cell and Developmental Biology - Professor of Biological Chemistry
Person: Academic