Functional characterization of the Drosophila suzukii pro-apoptotic genes reaper , head involution defective and grim
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Functional characterization of the Drosophila suzukii pro‑apoptotic genes reaper, head involution defective and grim Syeda A. Jaffri1 · Ying Yan1 · Jonas Schwirz2 · Marc F. Schetelig1,2 Accepted: 18 October 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Apoptosis is a fundamental process for the elimination of damaged or unwanted cells, and is a key aspect of development. It is triggered by pro-apoptotic genes responding to the intrinsic pathway that senses cell stress or the extrinsic pathway that responds to signals from other cells. The disruption of these genes can therefore lead to developmental defects and disease. Pro-apoptotic genes have been studied in detail in the fruit fly Drosophila melanogaster, a widely-used developmental model. However, little is known about the corresponding genes in its relative D. suzukii, a pest of soft fruit crops that originates from Asia but is now an invasive species in many other regions. The characterization of D. suzukii pro-apoptotic genes could lead to the development of transgenic sexing strains for pest management. Here, we describe the isolation and characterization of the pro-apoptotic genes reaper (Dsrpr), head involution defective (Dshid) and grim (Dsgrim) from a laboratory strain of D. suzukii. We determined their expression profiles during development, revealing that all three genes are expressed throughout development but Dsrpr is expressed most strongly, especially at the pupal stage. Functional analysis was carried out by expressing single genes or pairs (linked by a 2A peptide) in S2 cell death assays, indicating that Dsgrim and Dshid are more potent pro-apoptotic genes than Dsrpr, and the lethality can be significantly enhanced by co-expression of two genes. Therefore, the binary or multiple expression of different pro-apoptotic genes can be considered to build an efficient transgenic sexing system in D. suzukii. Keywords 2A peptide · RHG proteins · Sterile insect technique
Introduction
The GenBank accession numbers are as follows: Dshid mRNA: MN982930; Dsgrim mRNA: MN982931; and Dsrpr mRNA: MN982932. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10495-020-01640-2) contains supplementary material, which is available to authorized users. * Ying Yan [email protected]‑giessen.de * Marc F. Schetelig [email protected]‑giessen.de 1
Institute for Insect Biotechnology, Justus-Liebig-University Giessen, Winchesterstraße 2, 35394 Giessen, Germany
Fraunhofer IME-BR, Winchesterstraße 2, 35394 Giessen, Germany
2
Apoptosis (a form of programmed cell death) is an evolutionarily conserved process that eliminates unwanted cells during development as well as cells damaged by stress [1]. Apoptotic cells are characterized by plasma membrane blebbing, cytoplasmic condensation and shrinkage, progressive chromatin degradation, and nuclear fragmentation, before the apoptotic bodies are engulfed by cells of the immune system. The process can be triggered by intracellular
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