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PAP1 activates the prophenoloxidase system against bacterial infection in Musca domestica

Xu-Na Zhuang1, Yuan-Yuan Luan1, Tong-Rui Lv1

  • 1Biotechnology Department, School of Biological Sciences and Biotechnology, University of Jinan, Jinan, 250022, China.

Developmental and Comparative Immunology
|June 25, 2021
PubMed

Insights

The housefly prophenoloxidase-activating proteinase 1 (mdPAP1) gene is crucial for immunity. Its protein product activates phenoloxidase, enhancing the housefly

Area of Science:

  • Insect immunology
  • Molecular biology
  • Biochemistry

Background:

  • The housefly, Musca domestica, possesses a complex immune system.
  • Prophenoloxidase-activating proteinases (PAP) are key components in insect immune responses.
  • Three putative PAP genes (mdPAP1, mdPAP2, mdPAP3) were previously identified in M. domestica.

Purpose of the Study:

  • To clone and functionally characterize the mdPAP1 gene and its encoded protein.
  • To investigate the role of mdPAP1 in the prophenoloxidase activation cascade.
  • To determine the involvement of mdPAP1 in the housefly's defense against bacterial infections.

Main Methods:

  • cDNA cloning and sequence analysis of mdPAP1.
  • Recombinant protein expression and enzymatic activity assays.
  • Gene expression analysis using quantitative methods.
  • RNA interference (RNAi) for gene silencing.
  • Larval mortality assays.

Main Results:

  • The mdPAP1 cDNA is 1358 bp, encoding a 373-amino acid protein with a predicted molecular weight of 41.27 kDa and an isoelectric point of 6.25.
  • MdPAP1 expression was detected in all life stages and significantly upregulated in larvae challenged with bacteria.
  • Recombinant MdPAP1 effectively cleaved prophenoloxidase to phenoloxidase, increasing total phenoloxidase activity.
  • Gene silencing of mdPAP1 resulted in significantly increased larval mortality.

Conclusions:

  • mdPAP1 plays a vital role in the prophenoloxidase-activating system in houseflies.
  • mdPAP1 is essential for effective immune defense against bacterial infections in M. domestica.
  • The findings highlight mdPAP1 as a potential target for controlling housefly populations.
Keywords:
Antibacterial defenseMusca domesticaProphenoloxidase-activating proteinase 1 (PAP1)proPO activation

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