Abstract
Ubiquitination is a major post-translational modification involved in nearly all aspects of eukaryotic biology. Previous RNA-Seq studies showed that ubiquitination plays essential roles in the heat tolerance of Saccharina japonica, but to date, large-scale profiling of the ubiquitome in S. japonica has not been reported. To better understand the regulatory roles of ubiquitination in heat responses of S. japonica, we investigated its ubiquitome under normal and heat stress by the combination of affinity enrichment and high-resolution liquid chromatography-tandem mass spectroscopy analysis. Altogether, 3305 lysine ubiquitination sites in 1562 protein groups were identified. After normalization, 152 lysine ubiquitination sites in 106 proteins were significantly upregulated and 208 lysine ubiquitination sites in 131 proteins were significantly downregulated in response to heat stress. Protein annotation and functional analysis suggested that ubiquitination modulates a variety of essential cellular and physiological processes, including but not limited to the ubiquitin-26S proteasome system, ribosome, carbohydrate metabolism, and oxidative phosphorylation. Our results provide a global view of the heat response ubiquitome in S. japonica, and could facilitate future studies on the physiological roles of these ubiquitination-related proteins.
Highlights
Saccharina japonica is one of the most important aquacultured brown algae with world’s annual yield of eleven million tons, and has significant economic, social, and ecological values (FAO, http://www.fao.org/)
This study provides a global view of the function of ubiquitination in heat response of S. japonica, paving the ways for illustrating the ubiquitin-driven regulatory mechanism and shedding light on the improvement of heat tolerance in S. japonica
To test the relative abundance of ubiquitinated proteins under normal temperature and heat stress, proteins extracted from normal temperature and heat stress treatments were subjected to Western blot analysis with an anti-ubiquitin antibody
Summary
Saccharina japonica is one of the most important aquacultured brown algae with world’s annual yield of eleven million tons, and has significant economic, social, and ecological values (FAO, http://www.fao.org/). S. japonica is a kind of cold-temper kelp, native to the waters near Hokkaido, Japan. Comparing with the origin place, the water temperature of cultivation zone is higher, so the kelp is always subjected to heat stress [1]. As global temperatures get higher, the growth period of S. japonica is becoming shorter and the disease risk during the seedling breeding and cultivation increases, which will limit the further improvement of the yield and quality of this kelp [2]. It is significant to breed heat resistance varieties for the healthy and sustainable development of S. japonica aquaculture. It is necessary to elucidate the heat responsive mechanism and identify more functional genes which are involved in the heat tolerance in S. japonica
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