Two Roles for the Tenebrio molitor Relish in the Regulation of Antimicrobial Peptides and Autophagy-Related Genes in Response to Listeria monocytogenes.

Maryam Keshavarz,Yong Hun Jo,Tariku Tesfaye Edosa,Yeon Soo Han

doi:10.3390/insects11030188

Abstract

Relish is a key NF-κB transcription factor of the immune-deficiency (Imd) pathway that combats infection by regulating antimicrobial peptides (AMPs). Understanding of the fundamental role of Tenebrio molitor Relish (TmRelish) in controlling of Listeria monocytogenes virulence through the regulation of both AMPs and autophagy-related (ATG) genes is unclear. Here, we show that TmRelish transcripts were highly abundant in the larval fat body and hemocytes compared to the gut upon L. monocytogenes infection. Furthermore, significant mortality was observed in TmRelish-silenced larvae after intracellular insult. To investigate the cause of this lethality, we measured the induction of AMPs and ATG genes in the TmRelish dsRNA-treated T. molitor larvae. The expression of TmTenecin-1, TmTenecin-4, TmColeptericin-1, TmAttacin-2, and TmCecropin-2 were suppressed in the fat body and hemocytes of dsTmRelish-injected larvae during L. monocytogenes infection. In addition, TmRelish knockdown led to a noticeable downregulation of TmATG1 (a serine-threonine protein kinase) in the fat body and hemocytes of young larvae 6 h post-infection (pi). The notable increase of autophagy genes in the early stage of infection (6 h pi), suggesting autophagy response is crucial for Listeria clearance. Taken together, these results suggest that TmRelish plays pivotal roles in not only regulation of AMP genes but also induction of autophagy genes in response to L. monocytogenes challenge in fat body and hemocytes of T. molitor larvae. Furthermore, negative regulation of several AMPs by TmRelish in the fat body, hemocytes, and gut leaves open the possibility of a crosstalk between Toll and Imd pathway.

Highlights

The innate immune system is an evolutionarily conserved defense mechanism that combats infectious pathogens
Previous studies of the immune function of TmRelish have shown that this transcription factor can be detected in all examined tissues, with high levels of expression in the gut, fat body, and hemocytes, and relatively low levels of expression in Malpighian tubules and integument [32]
In hemocytes of L. monocytogenes-challenged larvae, the transcriptional levels of TmRelish reached their maximum at 6 h and gradually decreased (Figure 1B)

Summary

Introduction

The innate immune system is an evolutionarily conserved defense mechanism that combats infectious pathogens. Once pathogens breach the first line of defense, the sensors, peptidoglycan-recognition proteins (PGRPs), recognize their respective bacterial peptidoglycans (PGNs) [1]. The signal generated from the PGRP/PGN interaction involves other intracellular components of the Toll and immune deficiency (Imd) signaling pathways, and it triggers nuclear translocation of nuclear factor kappa-light-chain-enhancer (NF-κB) transcription factors of those pathways, such as Dorsal and Relish, respectively [2,3]. This signal transduction leads to the transcriptional expression of immune-related effectors, such as antimicrobial peptides (AMPs) [4]. PGRP-LE is capable of activation of Imd pathway, and prophenoloxidase (proPO) cascade [7]

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Discussion

Conclusion