Donovan Adriana, Brownlie Alison, Dorschner Michael O, Zhou Yi, Pratt Stephen J, Paw Barry H, Phillips Ruth B, Thisse Christine, Thisse Bernard, Zon Leonard I
Department of Hematology/Oncology, and Howard Hughes Medical Institute, Children's Hospital, Boston, MA 02115, USA.
Blood. 2002 Dec 15;100(13):4655-9. doi: 10.1182/blood-2002-04-1169. Epub 2002 Aug 22.
Iron is an essential nutrient required for the function of all cells, most notably for the production of hemoglobin in red blood cells. Defects in the mechanisms of iron absorption, storage, or utilization can lead to disorders of iron-limited erythropoiesis or iron overload. In an effort to further understand these processes, we have used the zebrafish as a genetic system to study vertebrate iron metabolism. Here we characterized the phenotype of chardonnay (cdy), a zebrafish mutant with hypochromic, microcytic anemia, and positioned the mutant gene on linkage group 11. The cdy gene was isolated by a functional genomics approach in which we used a combination of expression studies, sequence analyses, and radiation hybrid panel mapping. We identified erythroid-specific genes using a whole embryo mRNA in situ hybridization screen and placed these genes on the zebrafish genomic map. One of these genes encoded the iron transporter divalent metal transporter 1 (DMT1) and colocalized with the cdy gene. We identified a nonsense mutation in the cdy allele and demonstrated that, whereas wild-type zebrafish DMT1 protein can transport iron, the truncated protein expressed in cdy mutants is not functional. Our studies further demonstrate the conservation of iron metabolism in vertebrates and suggest the existence of an alternative pathway of intestinal and red blood cell iron uptake.
铁是所有细胞功能所必需的营养素,对红细胞中血红蛋白的产生尤为重要。铁吸收、储存或利用机制的缺陷可导致铁限制红细胞生成或铁过载紊乱。为了进一步了解这些过程,我们利用斑马鱼作为遗传系统来研究脊椎动物的铁代谢。在此,我们描述了霞多丽(cdy)的表型,这是一种患有低色素性、小细胞性贫血的斑马鱼突变体,并将突变基因定位在第11连锁群上。通过功能基因组学方法分离出cdy基因,我们在该方法中结合了表达研究、序列分析和辐射杂种板作图。我们使用全胚胎mRNA原位杂交筛选鉴定了红系特异性基因,并将这些基因定位在斑马鱼基因组图谱上。其中一个基因编码铁转运蛋白二价金属转运蛋白1(DMT1),并与cdy基因共定位。我们在cdy等位基因中鉴定出一个无义突变,并证明野生型斑马鱼DMT1蛋白可以转运铁,而cdy突变体中表达的截短蛋白没有功能。我们的研究进一步证明了脊椎动物中铁代谢的保守性,并提示存在肠道和红细胞铁摄取的替代途径。