V-ATPase subunit B plays essential roles in the molting process of the Chinese mitten crab, Eriocheir sinensis.

Xin Hou,Xiaowen Chen,Wucheng Yue,Hua Han,He Yang,Chenghui Wang,Jun Wang

doi:10.1242/bio.048926

Abstract

ABSTRACTVacuolar ATPase (V-ATPase) is a proton pump driven by ATP hydrolysis, and it plays an important role in numerous biological processes, such as protein degradation and synthesis, cell growth, and cell autophagy. The V-ATPase subunit B (VATB) is a conservative and regulatory subunit required for ATP hydrolysis during proton pumping. The VATB of Eriocheir sinensis (EsVATB), which includes an open reading frame (ORF) length of 1467 bp encoding 489 amino acids, was cloned to unveil the biological function of VATB during the molting process of crustaceans. Spatial and temporal expression profiles showed that EsVATB was highly expressed in the posterior gill accompanied with the highest osmotic pressure in the premolt (PrM) stage. Meanwhile, the highest expression level of EsVATB was identified in the hepatopancreas and heart during the postmolt stage and epidermis in the intermolt stage, indicating that EsVATB may perform diverse biological functions in different tissues during the molting process. The individual crabs in the interference group showed a high mortality rate (74%) and a low molting rate (26%) and failed to form a new epicuticle in the PrM stage. Meanwhile, a significant difference in osmotic pressure was identified between the interference and control groups. Our results indicate that EsVATB is an indispensable functional gene that may participate in osmoregulation and help with the new epicuticle formation during the molting process of E. sinensis.

Highlights

Vacuolar ATPase (V-ATPase) is one of the major classes of ATPase in eukaryotes, coupling the energy released from ATP hydrolysis to proton transport (Forgac, 1989; Harvey, 1992; Wieczorek et al, 1999)
BLASTP search by the deduced amino acid sequence of EsVATB revealed the highly conserved structure of this protein compared with other species
The two isoforms identified in this study may exhibit different expression regulation patterns in E. sinensis

Summary

Introduction

Vacuolar ATPase (V-ATPase) is one of the major classes of ATPase in eukaryotes, coupling the energy released from ATP hydrolysis to proton transport (Forgac, 1989; Harvey, 1992; Wieczorek et al, 1999). V-ATPase is essential in regulating physiological processes, such as endocytosis, protein degradation, amino acid transportation, uptake of neurotransmitters, intracellular pH homeostasis and waste disposal in the cell through numerous cell signaling pathways The structure of V-ATPase is conserved in invertebrate and vertebrate species, and studies have been conducted to elucidate its essential roles in biological processes in numerous organisms (McGuire et al, 2017)

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