Abstract
The larvae of the African midge, Polypedilum vanderplanki, can enter an ametabolic state called anhydrobiosis to overcome fatal desiccation stress. The Pv11 cell line, derived from P. vanderplanki embryo, shows desiccation tolerance when treated with trehalose before desiccation and resumes proliferation after rehydration. However, the molecular mechanisms of this desiccation tolerance remain unknown. Here, we performed high-throughput CAGE-seq of mRNA and a differentially expressed gene analysis in trehalose-treated, desiccated, and rehydrated Pv11 cells, followed by gene ontology analysis of the identified differentially expressed genes. We detected differentially expressed genes after trehalose treatment involved in various stress responses, detoxification of harmful chemicals, and regulation of oxidoreduction that were upregulated. In the desiccation phase, L-isoaspartyl methyltransferase and heat shock proteins were upregulated and ribosomal proteins were downregulated. Analysis of differentially expressed genes during rehydration supported the notion that homologous recombination, nucleotide excision repair, and non-homologous recombination were involved in the recovery process. This study provides initial insights into the molecular mechanisms underlying the extreme desiccation tolerance of Pv11 cells.
Highlights
Desiccation stress, the loss of essential water, can be fatal
To detect genes potentially related to desiccation tolerance and successful recovery after rehydration, we analysed differentially expressed gene (DEG) in control Pv11 cells (T0) and at different stages of anhydrobiosis: trehalose treatment for 48 h (T48), desiccation for 8 h or 10 days (D8 and D10d), and rehydration of D10d cells for 3 or 24 h (R3, R24)
Fewer genes encoding late embryogenesis abundant proteins (LEAs), TRXs, and protein L-isoaspartyl methyltransferase (PIMT) were significantly upregulated in Pv11 cells compared with in P. vanderplanki larvae
Summary
Desiccation stress, the loss of essential water, can be fatal. To tolerate desiccation stress, various organisms, such as rotifers, tardigrades, nematodes, plants, and larvae of the African midge Polypedilum vanderplanki, enter an ametabolic state called anhydrobiosis[1,2] and survive even if more than 99% of body water is lost[3]. The genome of P. vanderplanki has a paralogous gene cluster for TRXs8. These TRXs are upregulated by dehydration, and P. vanderplanki becomes tolerant to ROS-induced damage[8]. Desiccation tolerance of Pv11 cells is induced by treatment with culture medium containing 600 mM trehalose for 48 h. Analysis of gene expression patterns of Pv11 cells during trehalose treatment, desiccation, and rehydration may help to elucidate which genes are required to avoid cell death and return Pv11 cells from anhydrobiosis to the normal state. We performed high-throughput CAGE-seq of mRNA and a differentially expressed gene (DEG) analysis in trehalose-treated, desiccated, and rehydrated Pv11 cells, followed by gene ontology (GO) analysis of the identified DEGs. To the best of our knowledge, this is the first report of a comprehensive DEG analysis using CAGE-seq data for Pv11 cells that infers a putative mechanism of their avoidance of cell death and recovery from anhydrobiosis
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