Haploinsufficiency screen highlights two distinct groups of ribosomal protein genes essential for embryonic stem cell fate
Haploinsufficiency screen highlights two distinct groups of ribosomal protein genes essential for embryonic stem cell fate
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DOI：
10.1073/pnas.1418845112
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发表时间：
2015-02-17
影响因子：
11.1
通讯作者：
Sauvageau, Guy
中科院分区：
文献类型：
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作者：
Fortier, Simon;MacRae, Tara;Sauvageau, Guy
在对嵌套半合子染色体缺失的小鼠胚胎干细胞（ESCs）进行的功能基因组学筛选中，我们发现核糖体蛋白（RP）基因是胚状体（EB）形成的最重要的单倍不足决定因素。三个RP基因（Rps5, Rps14，或Rps28）的半合子性与最深刻的表型相关，其特征是其对应蛋白与核糖体mRNA出口位点的接近程度。这种EB表型被BAC或cDNA互补完全挽救，而不是p53水平的降低，尽管这种降低对大多数其他非mrna退出位点蛋白对应的rp缺失克隆是有效的。rna测序研究进一步显示，与野生型相比，Rps5半合子的未分化ESCs中几种中胚层特异性基因的表达水平降低。总之，这些结果表明，RP基因剂量限制了小鼠ESCs的分化，而不是自我更新。他们还强调了这一过程背后的两个独立机制，其中一个与p53无关。
In a functional genomics screen of mouse embryonic stem cells (ESCs) with nested hemizygous chromosomal deletions, we reveal that ribosomal protein (RP) genes are the most significant haploinsufficient determinants for embryoid body (EB) formation. Hemizygocity for three RP genes (Rps5, Rps14, or Rps28), distinguished by the proximity of their corresponding protein to the ribosome's mRNA exit site, is associated with the most profound phenotype. This EB phenotype was fully rescued by BAC or cDNA complementation but not by the reduction of p53 levels, although such reduction was effective with most other RP-deleted clones corresponding to non-mRNA exit-site proteins. RNA-sequencing studies further revealed that undifferentiated ESCs hemizygous for Rps5 showed reduced expression levels of several mesoderm-specific genes as compared with wild-type counterparts. Together, these results reveal that RP gene dosage limits the differentiation, not the self-renewal, of mouse ESCs. They also highlight two separate mechanisms underlying this process, one of which is p53 independent.