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Publications

Journal Article Epublish

PubMed: 39804945 DOI: 10.1371/journal.pntd.0012777

Stage-specific expression and divergent functions of two insulinase-like proteases associated with host infectivity in Cryptosporidium.

PLoS Negl Trop Dis 19(1):e0012777

Authors: Yue Huang, Shifeng Pei, Xin Lv, Fuxian Yang, Xiaoqing Gong, Na Li, Yaqiong Guo, Yaoyu Feng, Lihua Xiao

Affiliations

  • State Key Laboratory for Animal Disease Control and Prevention, South China Agricultural University, Guangzhou, China.
  • State Key Laboratory for Animal Disease Control and Prevention, South China Agricultural University, Guangzhou, China. Guangdong Laboratory for Lingnan Modern Agriculture, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.

Abstract

BACKGROUND

The determinants of differences in host infectivity among Cryptosporidium species and subtypes are poorly understood. Results from recent comparative genomic studies suggest that gains and losses of multicopy subtelomeric genes encoding insulinase-like proteases (INS-19 and INS-20 in Cryptosporidium parvum and their orthologs in closely related species) may potentially contribute to these differences.

METHODOLOGY/PRINCIPAL FINDINGS

In this study, we investigated the expression and biological function of the INS-19 and INS-20 of C. parvum. CRISPR/Cas9 was used to endogenously tag both genes with the hemagglutinin epitope. Immunofluorescence analysis revealed that INS-19 and INS-20 are expressed at different developmental stages of the pathogen. Although knockout of either had no detectable effect on the in vitro growth of C. parvum, knockout of INS-20, deletion of its multiple domains, or mutation of the active motif in the functional domain reduced the intensity of C. parvum infection in IFN-γ knockout mice. Consistent with this, mice infected with the INS-20-deleted mutant had reduced intestinal damage and parasite burden.

CONCLUSIONS/SIGNIFICANCE

These results suggest that INS-19 and INS-20 have stage-specific expression with distinct biological functions, and that the presence of the INS-20 in zoonotic C. parvum contributes to its infectivity and fitness in mice.

MeSH

Genes, Tropical Medicine, Peptide Hydrolases, Insulysin