Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA 01003, USA.
BMC Genet. 2012 Oct 16;13:86. doi: 10.1186/1471-2156-13-86.
WC1 co-receptors belong to the scavenger receptor cysteine-rich (SRCR) superfamily and are encoded by a multi-gene family. Expression of particular WC1 genes defines functional subpopulations of WC1(+) γδ T cells. We have previously identified partial or complete genomic sequences for thirteen different WC1 genes through annotation of the bovine genome Btau_3.1 build. We also identified two WC1 cDNA sequences from other cattle that did not correspond to sequences in the Btau_3.1 build. Their absence in the Btau_3.1 build may have reflected gaps in the genome assembly or polymorphisms among animals. Since the response of γδ T cells to bacterial challenge is determined by WC1 gene expression, it was critical to understand whether individual cattle or breeds differ in the number of WC1 genes or display polymorphisms.
Real-time quantitative PCR using DNA from the animal whose genome was sequenced ("Dominette") and sixteen other animals representing ten breeds of cattle, showed that the number of genes coding for WC1 co-receptors is thirteen. The complete coding sequences of those thirteen WC1 genes is presented, including the correction of an error in the WC1-2 gene due to mis-assembly in the Btau_3.1 build. All other cDNA sequences were found to agree with the previous annotation of complete or partial WC1 genes. PCR amplification and sequencing of the most variable N-terminal SRCR domain (domain 1 which has the SRCR "a" pattern) of each of the thirteen WC1 genes showed that the sequences are highly conserved among individuals and breeds. Of 160 sequences of domain 1 from three breeds of cattle, no additional sequences beyond the thirteen described WC1 genes were found. Analysis of the complete WC1 cDNA sequences indicated that the thirteen WC1 genes code for three distinct WC1 molecular forms.
The bovine WC1 multi-gene family is composed of thirteen genes coding for three structural forms whose sequences are highly conserved among individual cattle and breeds. The sequence diversity necessary for WC1 genes to function as a multi-genic pattern recognition receptor array is encoded in the genome, rather than generated by recombinatorial diversity or hypermutation.
WC1 共受体属于清道夫受体半胱氨酸丰富 (SRCR) 超家族,由多基因家族编码。特定 WC1 基因的表达定义了 WC1(+)γδ T 细胞的功能亚群。我们之前通过注释牛基因组 Btau_3.1 构建版,鉴定了十三个不同 WC1 基因的部分或完整基因组序列。我们还从其他牛中鉴定了两个 WC1 cDNA 序列,它们与 Btau_3.1 构建版中的序列不对应。它们在 Btau_3.1 构建版中的缺失可能反映了基因组组装中的缺口或动物之间的多态性。由于 γδ T 细胞对细菌挑战的反应由 WC1 基因表达决定,因此了解单个牛或品种在 WC1 基因数量上是否存在差异或是否存在多态性至关重要。
使用来自测序动物(“Dominette”)和代表十个牛品种的十六个其他动物的 DNA 进行实时定量 PCR 显示,编码 WC1 共受体的基因数量为十三个。提出了这十三个 WC1 基因的完整编码序列,包括由于 Btau_3.1 构建版中的错误组装而导致的 WC1-2 基因的错误纠正。所有其他 cDNA 序列均与之前的完整或部分 WC1 基因注释一致。对十三个 WC1 基因的最可变 N 端 SRCR 结构域(具有 SRCR“a”模式的结构域 1)的每个基因进行 PCR 扩增和测序显示,这些序列在个体和品种之间高度保守。在来自三个牛品种的 160 个结构域 1 序列中,除了描述的十三个 WC1 基因之外,没有发现其他序列。对完整 WC1 cDNA 序列的分析表明,十三个 WC1 基因编码三种不同的 WC1 分子形式。
牛 WC1 多基因家族由十三个基因组成,这些基因编码三种结构形式,其序列在个体牛和品种之间高度保守。WC1 基因作为多基因模式识别受体阵列发挥作用所需的序列多样性编码在基因组中,而不是通过重组多样性或高突变产生。