Bombyx mori PAT4 gene inhibits BmNPV infection and replication through autophagy.

Abstract

Proton-assisted amino acid transporter 4 (PAT4) is a member of the solute carrier (SLC) 36 family, which mediates the transmembrane transport of amino acids and their derivatives. However, the function of PAT4 in Bombyx mori is not clear. In this study, BmPAT4 was cloned and identified using PCR technology. Its open reading frame (ORF) includes 1,395 bp, encoding 464 amino acid (Aa). Moreover, the sequence of BmPAT4 has the highest similarity with wild Bombyx.mandarina, Spodoptera frugiperda and Spodoptera litura, and it has ten transmembrane domains. BmPAT4 was localized in the cell membrane and expressed in all tissues of the silkworm. After Bombyx mori nuclear polyhedrosis virus (BmNPV) infection, the expression of BmPAT4 in midgut, hemolymph and fat body was significantly up-regulated. In addition, overexpression of BmPAT4 in BmN cells could significantly inhibit the proliferation of BmNPV, and the expression of several genes in autophagy pathway decreased significantly. On the contrary, down-regulation of BmPAT4 expression by RNA interference can promote BmNPV replication and proliferation, and the expression of key genes in autophagy pathway is significantly increased. This is the first time to report that BmPAT4 has antiviral effect in silkworm. Moreover, the silkworm activates BmTORC1 via BmPAT4, which inhibits autophagy in silkworm cells, resulting in a lack of energy and raw materials for BmNPV infection and replication in cells.