Defects of protein production in erythroid cells revealed in a zebrafish Diamond-Blackfan anemia model for mutation in RPS19.
Defects of protein production in erythroid cells revealed in a zebrafish Diamond-Blackfan anemia model for mutation in RPS19.
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DOI：
10.1038/cddis.2014.318
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发表时间：
2014-07-24
影响因子：
9
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中科院分区：
文献类型：
--
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钻石-布莱克凡贫血（DBA）是一种罕见的先天性红细胞再生障碍性贫血，典型地出现在婴儿早期的DBA患者。大约25%的患者携带核糖体蛋白（RP）S19基因突变;已报告RPS 24、RPS 17、RPL 35 A、RPL 11和RPL 5突变。核糖体蛋白缺乏如何导致DBA中红细胞特异性缺陷尚未得到很好的阐明。为了对DBA中的主要核糖体缺陷进行遗传建模，我们通过在斑马鱼中使用TALEN介导的基因靶向产生了rps 19无效突变体。该突变系的分子特征表明，rps 19缺陷再现了DBA的红系缺陷，包括缺乏成熟红细胞和p53激活。值得注意的是，我们发现rps 19突变体的珠蛋白的生产被显着抑制，然而，珠蛋白的转录水平增加或不受影响的rps 19突变体胚胎。珠蛋白基因的RNA/蛋白质水平的这种解离在另一个具有rpl 11缺陷的斑马鱼DBA模型中得到证实。使用在红系细胞中特异性表达mCherry的转基因斑马鱼，我们发现当rps 19或rpl 11突变时，红系细胞中的蛋白质产生减少。L-亮氨酸处理减轻了红系细胞中蛋白质产生的缺陷，并部分挽救了rps 19和rpl 11突变体中的贫血表型。对该模型的分析表明，红系细胞中蛋白质产生的减少可能有助于DBA的血液特异性表型。此外，新产生的rps 19斑马鱼突变体应该作为一个有用的动物模型，研究DBA。我们的体内研究结果可能为DBA的未来治疗策略提供线索。
Diamond–Blackfan anemia (DBA) is a rare congenital red cell aplasia that classically presents during early infancy in DBA patients. Approximately, 25% of patients carry a mutation in the ribosomal protein (RP) S19 gene; mutations in RPS24, RPS17, RPL35A, RPL11, and RPL5 have been reported. How ribosome protein deficiency causes defects specifically to red blood cells in DBA has not been well elucidated. To genetically model the predominant ribosome defect in DBA, we generated an rps19 null mutant through the use of TALEN-mediated gene targeting in zebrafish. Molecular characterization of this mutant line demonstrated that rps19 deficiency reproduced the erythroid defects of DBA, including a lack of mature red blood cells and p53 activation. Notably, we found that rps19 mutants' production of globin proteins was significantly inhibited; however, globin transcript level was either increased or unaffected in rps19 mutant embryos. This dissociation of RNA/protein levels of globin genes was confirmed in another zebrafish DBA model with defects in rpl11. Using transgenic zebrafish with specific expression of mCherry in erythroid cells, we showed that protein production in erythroid cells was decreased when either rps19 or rpl11 was mutated. L-Leucine treatment alleviated the defects of protein production in erythroid cells and partially rescued the anemic phenotype in both rps19 and rpl11 mutants. Analysis of this model suggests that the decreased protein production in erythroid cells likely contributes to the blood-specific phenotype of DBA. Furthermore, the newly generated rps19 zebrafish mutant should serve as a useful animal model to study DBA. Our in vivo findings may provide clues for the future therapy strategy for DBA.
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