Screening of proteins interacting with infectious hypodermal and hematopoietic necrosis virus in Pacific white shrimp...

Infectious hypodermal and hematopoietic necrosis virus (IHHNV) is a major pathogen that severely impacts the shrimp aquaculture industry, and has lead to significant economic losses. Understanding the molecular response mechanisms of shrimp against IHHNV infection is crucial for developing effective disease control strategies. In order to deeply explore the molecular mechanism of Litopenaeus vannamei responds to IHHNV infection, this study screened the shrimp proteins interacting with IHHNV via the yeast two-hybrid system. The proteins encoded by IHHNV, including the capsid protein (CP), non-structural protein 1 (NS1) and non-structural protein 2 (NS2), were used as bait proteins, respectively, to screen against yeast library. Totally 63, 90, and 35 positive clones were obtained for CP, NS1, and NS2, respectively. After removal of duplicate genes, 14, 14, and 11 positive candidate proteins were obtained, respectively. Subsequently, 3, 4, and 5 candidate proteins were selected for backcross verification, and fatty acid binding protein 1-B.1-like (FABP1-B.1-like), elongation factor 2-like isoform X2 (EF2L-X2), and insulin-like growth factor-binding protein-related protein 1 (IGFBP-rP1) interacted with CP. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), Phosphopyruvate hydrolase (PEP), and Alpha-glucosidase (α-Glu) interacted with NS1. FABP1-B.1-like, α-Glu, importin subunit alpha-1 (IMPα1), 40S ribosomal protein S20-like isoform X1 (RPS20-X1), and NPC intracellular cholesterol transporter 2-like (NPC2-like) interacted with NS2. These interacting proteins provide an important reference basis for analyzing the interactions between IHHNV and the host, as well as for revealing the molecular mechanisms regulating viral infection.