Nickerson John M, Frey Ruth A, Ciavatta Vincent T, Stenkamp Deborah L
Ophthalmology Department, Emory University, Atlanta, GA 30322, USA.
Mol Vis. 2006 Dec 9;12:1565-85.
The interphotoreceptor retinoid-binding protein (IRBP) gene possesses an unusual structure, encoding multiple Repeats, each consisting of about 300 amino acids. Our goals were to gain insight into the function of IRBP, and to test the current model for the evolution of IRBP, in which Repeats were replicated from a simpler ancestral gene.
We employed a bioinformatics approach to analyze IRBP loci in recently completed or near-complete genome sequences of several vertebrates and nonvertebrate chordates. IRBP gene expression in zebrafish was evaluated by reverse transcriptase PCR (RT-PCR) and in situ mRNA hybridizations with gene-specific probes.
Patterns of exons and introns in the IRBP genes of tetrapods were highly similar, as were predicted amino acid sequences and Repeat structures. IRBP gene structure in teleost fish was more variable, and we report a new gene structure for two species, the Japanese puffer fish (Takifugu rubripes) and the zebrafish (Danio rerio). These teleost genomes contain a two-gene IRBP locus arranged head-to-tail in which the first gene, Gene 1, is intronless and contains a single large exon encoding three complete Repeats. It is followed by a second gene, Gene 2, which corresponds to the previously reported gene consisting of two Repeats spread across four exons and three introns. Each of the two zebrafish genes is transcribed. Gene 2 is expressed in the photoreceptors and RPE, and Gene 1 is expressed in the inner nuclear layer and weakly in the ganglion cell layer.
The tetrapod IRBP gene structure is highly conserved while the teleost fish gene structure was a surprise: It appears to be a two-gene locus with distinct Repeat organization in each open reading frame. This gene structure and gene expression data are consistent with possible neofunctionalization or sub-function partitioning of Gene 1 and Gene 2 in the zebrafish. We suggest that the two-gene locus in teleost fish arose as a consequence of either the known whole genome duplication or single gene tandem duplication.
光感受器间类视黄醇结合蛋白(IRBP)基因具有不同寻常的结构,编码多个重复序列,每个重复序列约由300个氨基酸组成。我们的目标是深入了解IRBP的功能,并检验当前关于IRBP进化的模型,该模型认为重复序列是从一个更简单的祖先基因复制而来的。
我们采用生物信息学方法分析了几种脊椎动物和非脊椎动物脊索动物最近完成或接近完成的基因组序列中的IRBP基因座。通过逆转录聚合酶链反应(RT-PCR)和用基因特异性探针进行原位mRNA杂交来评估斑马鱼中IRBP基因的表达。
四足动物IRBP基因中外显子和内含子的模式高度相似,预测的氨基酸序列和重复结构也是如此。硬骨鱼中的IRBP基因结构更具变异性,我们报道了两种物种——日本河豚(红鳍东方鲀)和斑马鱼(斑马鱼)的新基因结构。这些硬骨鱼基因组包含一个首尾相连排列的双基因IRBP基因座,其中第一个基因,即基因1,没有内含子,包含一个单一的大外显子,编码三个完整的重复序列。接着是第二个基因,即基因2,它对应于先前报道的由分布在四个外显子和三个内含子中的两个重复序列组成的基因。斑马鱼的这两个基因都被转录。基因2在光感受器和视网膜色素上皮中表达,基因1在内核层中表达,在神经节细胞层中表达较弱。
四足动物IRBP基因结构高度保守,而硬骨鱼的基因结构令人惊讶:它似乎是一个双基因座,每个开放阅读框中具有不同的重复序列组织。这种基因结构和基因表达数据与斑马鱼中基因1和基因2可能的新功能化或亚功能划分一致。我们认为硬骨鱼中的双基因座是已知的全基因组复制或单基因串联复制的结果。