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NM23 Is a CP-Binding Protein Involved in Infectious Hypodermal and Hematopoietic Necrosis Virus Infection in Shrimp
Xiaotong Yin1, Xiaoshan Wang1, Hui Sun1
1College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
Abstract:
The aim of this study was to identify the putative host cell receptor for Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) CP in the gill membrane of L. vannamei. Putative CP binding partners were screened first using a 2-dimensional Virus Overlay Protein Blot Assay (VOPBA) to probe isolated gill membrane proteins using recombinant CP. Putative binding partners were identified using mass spectrometry. A Phage Display Random Dodecapeptide Library was used to screen for dodecapeptides and motifs that bound to CP. Finally, putative binding pairs were confirmed using GST(glutathione-S-transferase) pulldown assays. 2-Dimensional VOPBA identified NM23 as a putative binding partner for IHHNV CP. GST pulldown experiments confirmed the direct interaction of NM23 and IHHNV CP. The phage display library was used to identify six groups of dodecapeptides that bound to CP. From these peptides, three characteristic binding motifs were identified, SW*Y, SKWV, and PQR. Interestingly, the SW*Y motif was also found in NM23. We are the first to implicate NM23 in IHHNV infection and postulate that it may bind to the CP using the SW*Y motif, although this remains to be confirmed.
Insights
Researchers identified NM23 as a potential host cell receptor for Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) in shrimp. This interaction may involve the SW*Y motif, offering new insights into IHHNV infection mechanisms.
Area of Science:
- Aquatic animal virology
- Molecular biology
- Biochemistry
Background:
- Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) causes significant disease in shrimp aquaculture.
- Identifying host cell receptors is crucial for understanding viral entry and developing control strategies.
- The specific receptor for IHHNV CP in *L. vannamei* gills remains unidentified.
Purpose of the Study:
- To identify the putative host cell receptor for IHHNV CP in the gill membrane of *L. vannamei*.
- To characterize the molecular interactions between IHHNV CP and potential host binding partners.
Main Methods:
- 2-dimensional Virus Overlay Protein Blot Assay (VOPBA) using recombinant IHHNV CP.
- Mass spectrometry for identification of putative binding partners.
- Phage Display Random Dodecapeptide Library screening.
- GST (glutathione-S-transferase) pulldown assays for confirmation of direct interactions.
Main Results:
- NM23 was identified as a putative binding partner for IHHNV CP via 2D VOPBA.
- GST pulldown assays confirmed a direct interaction between NM23 and IHHNV CP.
- Phage display identified three binding motifs (SW*Y, SKWV, PQR), with SW*Y found in NM23.
Conclusions:
- NM23 is implicated as a potential host cell receptor for IHHNV CP in *L. vannamei*.
- The SW*Y motif in NM23 may mediate the binding to IHHNV CP.
- This study provides the first evidence linking NM23 to IHHNV infection.
