p53-Independent cell cycle and erythroid differentiation defects in murine embryonic stem cells haploinsufficient for Diamond Blackfan anemia-proteins: RPS19 versus RPL5.
p53-Independent cell cycle and erythroid differentiation defects in murine embryonic stem cells haploinsufficient for Diamond Blackfan anemia-proteins: RPS19 versus RPL5.
复制标题
DOI：
10.1371/journal.pone.0089098
复制
发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Liu JM
中科院分区：
文献类型：
--
作者：
Singh SA;Goldberg TA;Henson AL;Husain-Krautter S;Nihrane A;Blanc L;Ellis SR;Lipton JM;Liu JM
钻石黑扇贫血(DBA)是一种罕见的遗传性骨髓衰竭综合征，由核糖体蛋白单倍体功能不全引起。DBA表现出明显的表型变异，通常表现为红系发育不良，与非红系特征不太一致。核糖体装配失败激活的P53通路被认为是导致DBA红系衰竭的原因之一。我们研究了在核糖体蛋白基因Rps19或Rpl5中存在基因陷阱突变的小鼠胚胎干细胞(ES)。两个突变体都表现出核糖体蛋白单倍体不足和多聚体缺陷。Rps19突变型ES细胞的P53蛋白表达显著增加，而Rpl5突变型ES细胞的P53蛋白表达无明显增加。在这两个突变体中，类胚体的形成都减少了，但非特异性地被P53基因敲除所挽救。当类胚体进一步分化为原始的红系集落时，两个突变体的集落形成都显著减少，这再次被P53抑制所拯救。Rps19突变型ES细胞细胞周期分析正常，而Rpl5突变型ES细胞G2/M期明显延迟，且不受P53基因敲除的影响。一致的是，Rpl5突变的ES细胞在液体培养中比Rps19突变的细胞有更明显的生长缺陷。我们的结论是，我们的RPS19和RPL5单倍体不足的小鼠ES细胞中的缺陷不能用P53的稳定性来充分解释，因为P53基因敲除似乎增加了亲本细胞和突变细胞的生长和分化潜力。我们的研究表明，基因捕获小鼠ES细胞是研究DBA发病机制的有用工具。
Diamond Blackfan anemia (DBA) is a rare inherited bone marrow failure syndrome caused by ribosomal protein haploinsufficiency. DBA exhibits marked phenotypic variability, commonly presenting with erythroid hypoplasia, less consistently with non-erythroid features. The p53 pathway, activated by abortive ribosome assembly, is hypothesized to contribute to the erythroid failure of DBA. We studied murine embryonic stem (ES) cell lines harboring a gene trap mutation in a ribosomal protein gene, either Rps19 or Rpl5. Both mutants exhibited ribosomal protein haploinsufficiency and polysome defects. Rps19 mutant ES cells showed significant increase in p53 protein expression; however, there was no similar increase in the Rpl5 mutant cells. Embryoid body formation was diminished in both mutants but nonspecifically rescued by knockdown of p53. When embryoid bodies were further differentiated to primitive erythroid colonies, both mutants exhibited a marked reduction in colony formation, which was again nonspecifically rescued by p53 inhibition. Cell cycle analyses were normal in Rps19 mutant ES cells, but there was a significant delay in the G2/M phase in the Rpl5 mutant cells, which was unaffected by p53 knockdown. Concordantly, Rpl5 mutant ES cells had a more pronounced growth defect in liquid culture compared to the Rps19 mutant cells. We conclude that the defects in our RPS19 and RPL5 haploinsufficient mouse ES cells are not adequately explained by p53 stabilization, as p53 knockdown appears to increase the growth and differentiation potential of both parental and mutant cells. Our studies demonstrate that gene trap mouse ES cells are useful tools to study the pathogenesis of DBA.
登录
查看更多内容
影响因子：
7.7
作者：
Ganguli, G;Back, J;Wasylyk, B
通讯作者：
Wasylyk, B
影响因子：
20.3
作者：
Montel-Hagen, Amelie;Blanc, Lionel;Taylor, Naomi
通讯作者：
Taylor, Naomi
影响因子：
3.6
作者：
McGowan KA;Mason PJ
通讯作者：
Mason PJ
DOI：
10.1016/j.bbadis.2009.08.002
发表时间：
2009-10
影响因子：
6.2
作者：
Badhai, Jitendra;Frojmark, Anne-Sophie;Davey, Edward J.;Schuster, Jens;Dahl, Niklas
通讯作者：
Dahl, Niklas
影响因子：
9.8
作者：
Gazda, Hanna T.;Sheen, Mee Rie;Beggs, Alan H.
通讯作者：
Beggs, Alan H.