Abstract
B4GALT5, also known as β-1, 4 galactosyltransferase V, is one of the members of β-1, 4 galactosyltransferase gene (B4GALT) family, which was concerned with embryonic development, tumor generation, other malignant diseases. In this study, we firstly cloned porcine B4GALT (pB4GALT5) from porcine alveolar macrophages, and predicted the structural domain and function of seven porcine β-1, 4 galactosyltransferase (I–VII) based on transcriptome analysis of PRRSV infected cells. Additionally, the upregulated porcine B4GALT5 expression was detected from PRRSV infected porcine alveolar macrophage (PAM) cells. The PRRSV proliferation were slightly inhibited in overexpression of pB4GALT5 transfected cells, the interaction of B4GALT5 and GP5 of PRRSV was firstly be detected by Co-IP, and the co-location between B4GALT5 and GP5 were also observed in golgi membranes by confocal microscopy. A significant increasing mRNA transcription, including inflammatory cytokines (IFN-α, IL-6, IL-18, IL-1β, TNF-α) and some cell surface glycosylated protein involved in antigen present (MHC-I/II), cell adhesion and migration (chemokine MCP-1 and receptor CCR2; LFA-1, ICAM-1) were upregulated in B4GALT5 overexpressed PRRSV infected cells. Our results demonstrated that the regulation of pB4GALT5 plays an important roles in PRRSV proliferation and modification function in viral infection cells. And these results will make achievements by supporting the research of latent mechanisms of β-1, 4 galactosyltransferase V in antiviral immunity.
Highlights
B4GALT5, known as β-1, 4 galactosyltransferase V, is one of β-1, 4 galactosyltransferase (B4GALT) family including seven members (Zhou et al, 2013)
Current research has shown that β-1, 4 galactosyltransferases are concerned with embryonic development (Kumagai et al, 2009), Regulatory Roles of B4GALT5 in Porcine reproductive and respiratory syndrome virus (PRRSV) Multiplication neurological development (Shen et al, 2003a,b), immune and inflammatory responses (Parker et al, 2016), tumor generation and development (Shirane et al, 2014), and other life activities (Wei et al, 2010) and diseases (Cartault et al, 2015)
Porcine alveolar macrophages (PAM) cell line 3D4/21 grown in RPMI-1640 medium (Gibco) added with 10% fetal bovine serum (FBS) (Biological Industries)
Summary
B4GALT5, known as β-1, 4 galactosyltransferase V, is one of β-1, 4 galactosyltransferase (B4GALT) family including seven members (Zhou et al, 2013). B4GALT is one of the most studied glycosyltransferases in recent years. These enzymes catalyze the shift of galactosyl group from UDP galactoside to N-acetylglucosamine or other glycosyl acceptor to determine the biosynthesis of different glycoconjugates (Zhang et al, 2017). Substrate specificity and distribution in different tissues of the members of the β-1, 4 galactosyltransferase family determine their functional differences (Fan et al, 1999; Ujita et al, 2000). The functional aspect of β4Ga1T1 is the most studied, followed by β4Ga1T5
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