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Proc Natl Acad Sci U S A. 2015 Feb 17;112(7):2127-32. doi: 10.1073/pnas.1418845112. Epub 2015 Feb 2.

Haploinsufficiency screen highlights two distinct groups of ribosomal protein genes essential for embryonic stem cell fate.

Author information

1
Molecular Genetics of Stem Cells Laboratory, Institute for Research in Immunology and Cancer, University of Montreal, Montreal, QC, H3C 3J7, Canada;
2
Molecular Genetics of Stem Cells Laboratory, Institute for Research in Immunology and Cancer, University of Montreal, Montreal, QC, H3C 3J7, Canada; Division of Molecular Medicine, The Walter and Eliza Hall Institute of Medical Research, Melbourne, VIC 3050, Australia; and Department of Medical Biology, The University of Melbourne, Parkville, VIC 3010, Australia.
3
Molecular Genetics of Stem Cells Laboratory, Institute for Research in Immunology and Cancer, University of Montreal, Montreal, QC, H3C 3J7, Canada; guy.sauvageau@umontreal.ca.

Abstract

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.

KEYWORDS:

differentiation; embryoid body; embryonic stem cells; ribosomal proteins

PMID:
25646475
PMCID:
PMC4343140
DOI:
10.1073/pnas.1418845112
[Indexed for MEDLINE]
Free PMC Article

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