Duck Antibody Genes: Unusual Organization and Expression

点击翻译按钮获取中文摘要

The long-term goal of the proposed studies is to understand the genetic mechanisms controlling the expression of duck immunoglobulin genes. The duck has an immune response that can be characterized as ineffectual, which has significance for human health given the role of ducks (and other waterfowl) as carriers (or reservoirs) of diseases infectious for man. E.g., ducks constitute the only year-round reservoir of avian influenza A viruses, which are a source of genetic variation implicated in the emergence of novel influenza strains pathogenic to man. Understanding the molecular basis of the expression of duck antibodies will permit an understanding of their ineffectual antibody response and also contribute significantly to our understanding of the evolution of transcriptional control, an important aspect of comparative genomics. The duck has an unusual Ig heavy chain (IgH) locus, which has undergone significant rearrangement in its germline organization. The 3 constant (C) region genes are arranged mu-alpha-upsilon, with the alpha gene in reverse transcriptional orientation. This leads to unique problems of transcriptional control associated particularly with the control of class switch recombination. These are addressed in the Specific Aims of this proposal, and the hypotheses to be tested can be phrased as 3 questions: 1) Does the alpha gene have its own I-exon or does it rely on the bi-directional I-upsilon promoter to initiate class switching to IgA? 2) Have the signal transduction pathways that control Ig class-switching, by transcription from I-exon promoters, been conserved in vertebrate evolution? 3) The mammalian IgH locus contains transcriptional enhancers in both the JH to mu intron and 3' of the C-gene cluster. Where, in the duck IgH locus are the enhancers, what transcription factor-binding motifs do they contain, and how do they function? The approach involves identifying transcriptional control elements through functional analysis (i.e. their ability to drive transcription of a reporter gene), and the responsiveness of I-exon promoters to cytokine-induced class-switch signals. The specific motifs involved will be identified by site-directed mutagenesis, and the transcription factors binding these sites will be identified through EMSA/antibody supershift approaches.The results of these proposed studies will enhance our understanding of the molecular basis of the unusual antibody response of the duck, and provide insight into the evolution of transcriptional control in the vertebrate immune system.