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Updated: May 24, 2025
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Published on: June 29, 2018
Proteomic profiling of kars knockout zebrafish larvae
Jingjing Wang1, Xiao Yu1, Ying Wang1
1Department of Otolaryngology-Head and Neck Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai 200233, China; Otolaryngology Institute of Shanghai Jiao Tong University, Shanghai 200233, China.
Background:
KARS encodes both mitochondrial and cytoplasmic lysyl-tRNA synthetase, which is one of the aminoacyl-tRNA synthetases (ARSs) necessary for protein translation. Pathogenic variants in KARS have been reported to be involved in hearing loss, visual disorders, neuropathology, and diseases combined with multisystem phenotypes. In vitro studies have shown that KARS mutations cause a decrease in aminoacylation. However, the pathogenetic mechanisms underlying the complex neurological phenotypes remain largely unknown.
Methods:
We developed kars knockout zebrafish and proteomic analyses on larvae with different genotypes at five days post-fertilization were performed using isobaric tags for relative and absolute quantitation (iTRAQ). Then the differentially abundant proteins (DAPs) analyzed by iTRAQ were validated by parallel reaction monitoring (PRM).
Results:
420 differentially abundant proteins were identified between the knockout and wildtype groups, of which, 138 were up-regulated and 282 down-regulated. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analyses showed the greatest DAP cluster enrichment in ribosome (P = 2.1 × 10-6, 28 genes), aminoacyl-tRNA biosynthesis (P = 7.34 × 10-6, 13 genes), and hypertrophic cardiomyopathy (P = 7.45 × 10-6, 28 genes). A further PRM-based analysis identified changes in nars, mybphb, atp2a1l, col6a1 and rps3a that were specially linked to kars-deficency.
Conclusions:
This work provides new valuable in vivo data for understanding the molecular mechanism of KARS deficiency-associated diseases, and will give us comprehensive insights into ARS-related disorders.
