Abstract
Alternative splicing is an essential molecular mechanism that increase the protein diversity of a species to regulate important biological processes. Ecdysone receptor (EcR), an essential nuclear receptor, is essential in the molting, growth, development, reproduction, and regeneration of crustaceans. In this study, the whole sequence of EcR gene from Eriocheir sinensis was obtained. The sequence was 45,481 bp in length with 9 exons. Moreover, four alternatively spliced EcR isoforms (Es-EcR-1, Es-EcR-2, Es-EcR-3 and Es-EcR-4) were identified. The four isoforms harbored a common A/B domain and a DNA-binding region but different D domains and ligand-binding regions. Three alternative splicing patterns (alternative 5′ splice site, exon skipping, and intron retention) were identified in the four isoforms. Functional studies indicated that the four isoforms have specific functions. Es-EcR-3 may play essential roles in regulating periodic molting. Es-EcR-2 may participate in the regulation of ovarian development. Our results indicated that Es-EcR has broad regulatory functions in molting and development and established the molecular basis for the investigation of ecdysteroid signaling related pathways in E. sinensis.
Highlights
Ecdysteroids are crucial to the growth, reproduction, development, regeneration and molting of crustaceans[1,2]
Based on the assembled draft genome of E. sinensis, we identified and evaluated the complete gene structures and preliminary functions of the four Ecdysone receptor (EcR) isoforms
These findings indicate that the crustacean EcR gene has a different selection pressures and evolutionary trajectories compared to insects (Table 1, Fig. 3)
Summary
Ecdysteroids are crucial to the growth, reproduction, development, regeneration and molting of crustaceans[1,2]. Alternative splicing is tightly regulated in a manner that is specific to cell or developmental stage and is a common biological process in human and other organisms[13]. Alternative spliced EcR isoforms have been identified and studied in both insects and crustaceans[6,17]. Alternative EcR isoforms have been identified in other insect species with specific functions in different tissues and developmental stages[23,24,25,26,27,28,29,30]. The crustaceans EcR gene has similar functions to insects, but has different numbers and structures of isoforms[6]. To thoroughly understand the biological function of the different isoforms in specific molting stages and tissues, semi-quantitative, quantitative gene expression analyses, and RNA interference experiments were conducted during molting and different tissues of E. sinensis
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