4.6 Article

Clip-Domain Serine Protease Gene (LsCLIP3) Is Essential for Larval-Pupal Molting and Immunity in Lasioderma serricorne

Journal

FRONTIERS IN PHYSIOLOGY
Volume 10, Issue -, Pages -

Publisher

FRONTIERS MEDIA SA
DOI: 10.3389/fphys.2019.01631

Keywords

clip-domain serine protease; Lasioderma serricorne; RNA interference; molting; immunity

Categories

Funding

  1. National Natural Science Foundation of China [31501649]
  2. Program for Science and Technology Outstanding talents in Higher Learning Institution of Guizhou Province [2017083]
  3. Program for Science and Technology Youth Talents in Department of Education in Guizhou Province [2018298]
  4. Discipline and Master's Site Construction Project of Guiyang University by Guiyang City Financial Support Guiyang University [SH-2019]
  5. Program for First-class Discipline Construction in Guizhou Province [201785]

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Clip-domain serine proteases (CLIPs) play crucial roles in insect development and innate immunity. In this study, we identified a CLIP gene (designated LsCLIP3) from the cigarette beetle Lasioderma serricorne. LsCLIP3 contains a 1,773-bp open reading frame (ORF) encoding a 390-amino-acid protein and shows a conserved clip domain and a trypsin-like serine protease domain. Phylogenetic analysis indicated that LsCLIP3 was orthologous to the CLIP-B subfamily. LsCLIP3 was prominently expressed in larva, pupa, and early adult stages. In larval tissues, it was highly expressed in the integument and fat body. The expression of LsCLIP3 was induced by 20-hydroxyecdysone. A similar induction was also found by peptidoglycans from Escherichia coli and Staphylococcus aureus. RNA interference (RNAi)-mediated silencing of LsCLIP3 disrupted larval-pupal molting and specifically reduced the expression of genes in 20-hydroxyecdysone synthesis and signaling pathway. The chitin amounts of LsCLIP3 RNAi larvae were greatly decreased, and expressions of six chitin metabolic-related genes were significantly reduced. Knockdown of LsCLIP3 increased larval sensitivity to Gram-negative and Gram-positive bacteria. There was significantly decreased expression of four antimicrobial peptide (AMP) genes. The results suggest that LsCLIP3 is an important component of the larva to pupa molt and for the immunity of L. serricorne.

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