The Penaeus stylirostris densovirus capsid protein interacts with the Litopenaeus vannamei BCCIP protein

Fish Shellfish Immunol. 2019 May:88:198-206. doi: 10.1016/j.fsi.2019.02.057. Epub 2019 Mar 1.

Abstract

Viral capsid proteins play an important role in the viral infection process. To identify the cellular proteins in shrimp that interact with the Penaeus stylirostris densovirus capsid protein (PstDNV-CP), we constructed a yeast two-hybrid (Y2H) cDNA library of the muscle tissue of Litopenaeus vannamei, and hybridized the bait vector pGBKT7-CP with this library. Cloning and sequencing showed that the shrimp protein interacting with PstDNV-CP was a homolog of BRCA2 and CDKN1A(p21)-interacting protein (BCCIP). We named this protein L. vannamei BCCIP (LvBCCIP). Further analysis showed that LvBCCIP interacted with L. vannamei calmodulin (LvCaM). We validated the interactions between PstDNV-CP and LvBCCIP, and between LvBCCIP and LvCaM, with GST pulldown assays. The gene expression of LvBCCIP increased significantly after PstDNV challenge. In addition, the PstDNV titer of PstDNV-challenged shrimp was significantly reduced after LvBCCIP expression was inhibited using double-stranded RNA (dsRNA) interference. These results indicated that LvBCCIP is critical to PstDNV pathogenesis in L. vannamei. Interestingly, the growth rate of L. vannamei was significantly reduced when LvBCCIP gene expression was silenced, indicating that LvBCCIP may also be associated with growth regulation in L. vannamei. Thus, the interaction between PstDNV-CP and LvBCCIP might explain why PstDNV infection leads to runt-deformity syndrome in shrimp.

Keywords: BCCIP; Capsid protein; Litopenaeus vannamei; Penaeus stylirostris densovirus.

MeSH terms

  • Animals
  • BRCA2 Protein / metabolism
  • Calmodulin / metabolism
  • Capsid Proteins / metabolism*
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism
  • Densovirus / physiology*
  • Gene Expression
  • Penaeidae / growth & development
  • Penaeidae / virology*
  • RNA Interference

Substances

  • BRCA2 Protein
  • Calmodulin
  • Capsid Proteins
  • Cyclin-Dependent Kinase Inhibitor p21