The histone H2A deubiquitinase Usp16 regulates embryonic stem cell gene expression and lineage commitment.
Polycomb Repressive Complex 1 and histone H2A ubiquitination (ubH2A) contribute to embryonic stem cell (ESC) pluripotency by repressing lineage-specific gene expression. However, whether active deubiquitination co-regulates ubH2A levels in ESCs and during differentiation is not known. Here we report that Usp16, a histone H2A deubiquitinase, regulates H2A deubiquitination and gene expression in ESCs, and importantly, is required for ESC differentiation. Usp16 knockout is embryonic lethal in mice, but does not affect ESC viability or identity. Usp16 binds to the promoter regions of a large number of genes in ESCs, and Usp16 binding is inversely correlated with ubH2A levels, and positively correlates with gene expression levels. Intriguingly, Usp16(-/-) ESCs fail to differentiate due to ubH2A-mediated repression of lineage-specific genes. Finally, Usp16, but not a catalytically inactive mutant, rescues the differentiation defects of Usp16(-/-) ESCs. Therefore, this study identifies Usp16 and H2A deubiquitination as critical regulators of ESC gene expression and differentiation.
Department of Biochemistry and Molecular Genetics, Stem Cell Institute, University of Alabama at Birmingham, Kaul Human Genetics Building 430, 720 South 20th Street, Birmingham, Alabama 35294, USA.
Wei
W
Yang
Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, 8 Medical Drive, MD7 #02-07, Singapore 117597, Singapore.
Yun-Hwa
YH
Lee
Department of Biochemistry and Molecular Genetics, Stem Cell Institute, University of Alabama at Birmingham, Kaul Human Genetics Building 430, 720 South 20th Street, Birmingham, Alabama 35294, USA.
Amanda E
AE
Jones
Department of Biochemistry and Molecular Genetics, Stem Cell Institute, University of Alabama at Birmingham, Shelby Building 770, 1825 University Boulevard, Birmingham, Alabama 35294, USA.
Jessica L
JL
Woolnough
Department of Biochemistry and Molecular Genetics, Stem Cell Institute, University of Alabama at Birmingham, Kaul Human Genetics Building 530, 720 South 20th Street, Birmingham, Alabama 35294, USA.
Dewang
D
Zhou
Department of Pathology, University of Alabama at Birmingham, West Pavilion, 619 South 19th Street, Birmingham, Alabama 35233, USA.
Qian
Q
Dai
Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, 8 Medical Drive, MD7 #02-07, Singapore 117597, Singapore.
Qiang
Q
Wu
Department of Biochemistry and Molecular Genetics, Stem Cell Institute, University of Alabama at Birmingham, Shelby Building 770, 1825 University Boulevard, Birmingham, Alabama 35294, USA.
Keith E
KE
Giles
Department of Biochemistry and Molecular Genetics, Stem Cell Institute, University of Alabama at Birmingham, Kaul Human Genetics Building 530, 720 South 20th Street, Birmingham, Alabama 35294, USA.
Tim M
TM
Townes
Department of Biochemistry and Molecular Genetics, Stem Cell Institute, University of Alabama at Birmingham, Kaul Human Genetics Building 430, 720 South 20th Street, Birmingham, Alabama 35294, USA.
Hengbin
H
Wang
D000818:Animals; D002454:Cell Differentiation; D019070:Cell Lineage; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D005804:Genes, Lethal; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D011485:Protein Binding; D043222:Ubiquitin Thiolesterase / Q000378:metabolism / Q000502:physiology*
C584021:Usp16 protein, mouse; D043222:Ubiquitin Thiolesterase