RESUMO
TRIM25, a member of the tripartite motif-containing (TRIM) family of proteins, plays an important role in cell proliferation, protein modification, and the RIG-I-mediated antiviral signaling pathway. However, relatively few studies have investigated the molecular characterization, tissue distribution, and potential function of TRIM25 in chickens. In this study, we cloned the full-length cDNA of chicken TRIM25 that is composed of 2706 bp. Sequence analyses revealed that TRIM25 contains a 1902-bp open-reading frame that probably encodes a 633-amino acid protein. Multiple comparisons with deduced amino acid sequences revealed that the RING finger and B30.2 domains of chicken TRIM25 share a high sequence similarity with human and murine TRIM25, indicating that these domains are critical for the function of chicken TRIM25. qPCR assays revealed that TRIM25 is highly expressed in the spleen, thymus and lungs in chickens. Furthermore, we observed that TRIM25 expression was significantly upregulated both in vitro and in vivo following infection with Newcastle disease virus. TRIM25 expression was also significantly upregulated in chicken embryo fibroblasts upon stimulation with poly(I:C) or poly(dA:dT). Taken together, these findings suggest that TRIM25 plays an important role in antiviral signaling pathways in chickens.
Assuntos
Proteínas Aviárias/genética , Galinhas/genética , Doença de Newcastle/genética , Fatores de Transcrição/genética , Ubiquitina-Proteína Ligases/genética , Dedos de Zinco/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Proliferação de Células/genética , Clonagem Molecular , Proteínas de Ligação a DNA/genética , Expressão Gênica , Humanos , Camundongos , Dados de Sequência Molecular , Doença de Newcastle/virologia , Vírus da Doença de Newcastle , Poli A/farmacologia , Poli I-C/farmacologia , Poli T/farmacologia , Receptores do Ácido Retinoico/metabolismo , Análise de Sequência de DNA , Homologia de Sequência , Transdução de SinaisRESUMO
Myeloid differentiation primary response gene 88 (MYD88), a universal adapter protein, plays an important role in activating the nuclear factor-κB (NF-κB) and regulating the expression of proinflammatory genes like tumor necrosis factor (TNF) and interleukin-1 (IL-1), which were highly involved in Salmonella Pullorum infection. To detect the relationship between polymorphisms of the MyD88 gene and Salmonella Pullorum disease, we screened the coding region (CDS) of the MYD88 gene by DNA pool construction and sequencing based on case-control study. Eight single nucleotide polymorphisms (SNPs) in the sequenced fragment (5 exons), 7 known loci and one novel mutation named G4810372T (SNP8), were found in the fifth exon. In addition, we found 7 nonsynonymous substitutions. The allele frequency of only one SNP, g.4810191C > T (SNP1), was significantly different (P < 0.05) between case and control groups. The genotype frequencies of SNP1 (g.4810191C > T) and SNP3 (g.4810257G > T) were of significant difference between the case and the control groups (P < 0.05). Collectively, SNPs of the MyD88 gene were significantly associated with susceptibility to Salmonella Pullorum infection, which can be used as a disease-resistant marker in chicken. These results provided a theoretical basis for future research on chicken breeding by marker-assisted selection.
Assuntos
Galinhas/genética , Predisposição Genética para Doença/genética , Salmonelose Animal/genética , Animais , Estudos de Casos e Controles , Haplótipos , Desequilíbrio de Ligação/genética , Polimorfismo de Nucleotídeo Único/genética , SalmonellaRESUMO
It has been known that the chicken's resistance to disease was affected by chicken's genetic background. And RLR-mediated antiviral pathway plays an important role in detection of viral RNA. However, little is known about the interaction of genetic background with RLR-mediated antiviral pathway in chicken against MDV infection. In this study, we adopted economic line-AA broilers and native Erlang mountainous chickens for being infected with MDV. Upon infection with MDV, the expression of MDA-5 was upregulated in two-breed chickens at 4, 7, and 21 d.p.i. It is indicated that MDA-5 might be involved in detecting MDV in chicken. Interestingly, the expression of IRF-3 and IFN- ß genes was decreased in spleen and thymus of broilers at 21 d.p.i, but it was upregulated in immune tissues of Erlang mountainous chickens. And the genome load of MDV in spleen of broiler is significantly higher than that in Erlang mountainous chickens. Meanwhile, we observed that the death of broiler mainly also occurred in this phase. Collectively, these present results demonstrated that the expression patters of IRF-3 and IFN- ß genes in chicken against MDV infection might be affected by the genetic background which sequently influence the resistance of chicken response to MDV.