Identification of a dengue 2 virus envelope protein receptor in Aedes aegypti critical for viral midgut infection

• 成果类型: Article
• 署名作者: Kantor, Asher M.; Talyuli, Octavio A. C.; Reid, William R.; Alvarenga, Patricia Hessab; Booker, Jasmine; Lin, Jingyi; Franz, Alexander W. E.; Barillas, Carolina
• 署名单位: National Institutes of Health (NIH) - USA; NIH National Institute of Allergy & Infectious Diseases (NIAID); University of Missouri System; University of Missouri Columbia
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-14847
• DOI: 10.1073/pnas.2417750121
• 发表日期: 2024-11-26
• 关键词: web tool; c6/36; glycoprotein; chopchop; immunity; domains; binding; cells; entry

摘要

The establishment of a productive dengue virus (DENV) infection in the midgut epithelial cells of Aedes aegypti is critical for the viral transmission cycle. The hypothesis that DENVvirions interact directly with specific mosquito midgut proteins was explored. We found that DENV serotype 2 (DENV2) pretreated with trypsin interacted with a single 31 kDa protein, identified as AAEL011180 by protein mass spectrometry. This putative receptor is a highly conserved protein and has orthologs in culicine and anopheline mosquitoes. We confirmed that impairing the expression of AAEL011180 in the midgut of Ae. aegypti females abolished the interaction with DENV2, and the virus also bound to immobilized recombinant purified receptor. Furthermore, recombinant DENV2 surface E glycoprotein bound to recombinant AAEL011180 with high affinity (38.2 nM) in binding kinetic analysis using surface plasmon resonance. The gene for this DENV2 E protein receptor (EPrRec) was disrupted, but since the gene is essential in Ae. aegypti, only heterozygote knockout (Delta EPrRec+/-) females could be recovered. Further reducing EPrRec mRNA expression in the midgut of Delta EPrRec +/- females by systemic dsRNA injection significantly reduced the prevalence of DENV2 midgut infection. EPrRec also interacts with heat shock protein 70 cognate 3 (Hsc70-3), and silencing Hsc70-3 expression in Delta EPrRec females also reduced the prevalence of DENV2 midgut infection.