Essential Role for Polycomb Group Protein Pcgf6 in Embryonic Stem Cell Maintenance and a Noncanonical Polycomb Repressive Complex 1 (PRC1) Integrity

Zhao, Wukui; Tong, Hanxing; Huang, Yikai; Yan, Yun; Teng, Huajian; Xia, Yin; Jiang, Qing; Qin, Jinzhong

doi:10.1074/jbc.m116.763961

Cited by 50 publications

(90 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, we found that deletion of Pcgf1 resulted in the reduction of Ring1B and its associated histone mark H2AK119ub1 at Pcgf1 target genes. This result is similar to those observed for Pcgf6 −/− ES cells, suggesting a common molecular mechanism controlling Polycomb recruitment by Pcgf family in ES cells43. Of note, the occupancy of BcoR (another component of PRC1.1) at these targets is also decreased.…”

Section: Discussionsupporting

confidence: 86%

Loss of Polycomb Group Protein Pcgf1 Severely Compromises Proper Differentiation of Embryonic Stem Cells

Yan

Zhao

Huang

et al. 2017

Sci Rep

Self Cite

View full text Add to dashboard Cite

The Polycomb repressive complex 1 (PRC1) is essential for fate decisions of embryonic stem (ES) cells. Emerging evidence suggests that six major variants of PRC1 complex, defined by the mutually exclusive presence of Pcgf subunit, regulate distinct biological processes, yet very little is known about the mechanism by which each version of PRC1 instructs and maintains cell fate. Here, we disrupted the Pcgf1, also known as Nspc1 and one of six Pcgf paralogs, in mouse ES cells by the CRISPR/Cas9 technology. We showed that although these mutant cells were viable and retained normal self-renewal, they displayed severe defects in differentiation in vitro. To gain a better understanding of the role of Pcgf1 in transcriptional control of differentiation, we analysed mRNA profiles from Pcgf1 deficient cells using RNA-seq. Interestingly, we found that Pcgf1 positively regulated expression of essential transcription factors involved in ectoderm and mesoderm differentiation, revealing an unexpected function of Pcgf1 in gene activation during ES cell lineage specification. Chromatin immunoprecipitation experiments demonstrated that Pcgf1 deletion caused a decrease in Ring1B and its associated H2AK119ub1 mark binding to target genes. Altogether, our results suggested an unexpected function of Pcgf1 in gene activation during ES cell maintenance.

show abstract

Section: Discussionsupporting

confidence: 86%

Loss of Polycomb Group Protein Pcgf1 Severely Compromises Proper Differentiation of Embryonic Stem Cells

Yan

Zhao

Huang

et al. 2017

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…S2 and S3). These observations, together with our previous findings (19,20), strongly suggest that all PCGF family members except for Pcgf6 are dispensable for the self-renewal of ES cells.…”

Section: Pcgf3/5 Are Not Essential For Es Cell Self-renewalsupporting

confidence: 81%

“…To delete the region encoding the RING finger motif and introduce a frameshift, we designed two single guide RNAs (sgRNAs) specifically flanking exon 2 of the mouse Pcgf3 or Pcgf5 gene. Independent ES cell clones were isolated, and genomic deletions were confirmed by PCR with primers outside the expected cleavage site (19) (Fig. S1).…”

Section: Pcgf3/5 Are Not Essential For Es Cell Self-renewalmentioning

confidence: 99%

Polycomb group RING finger proteins 3/5 activate transcription via an interaction with the pluripotency factor Tex10 in embryonic stem cells

Zhao

Huang

Zhang

et al. 2017

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Polycomb group (PcG) proteins are epigenetic transcriptional repressors that orchestrate numerous developmental processes and have been implicated in the maintenance of embryonic stem (ES) cell state. More recent evidence suggests that a subset of PcG proteins engages in transcriptional activation in some cellular contexts, but how this property is exerted remains largely unknown. Here, we generated ES cells with single or combined disruption of polycomb group RING finger protein 3 (Pcgf3) and Pcgf5 with the CRISPR-Cas9 technique. We report that although these mutant cells maintained their self-renewal and colony-forming capacity, they displayed severe defects in mesoderm differentiation and Using RNA-seq to analyze transcriptional profiles of ES cells with single or combined Pcgf3/5 deficiencies, we found that in contrast to the canonical role of the related polycomb repressive complex 1 (PRC1) in gene repression, Pcgf3/5 mainly function as transcriptional activators driving expression of many genes involved in mesoderm differentiation. Proteomic approaches and promoter occupancy analyses helped to establish an extended Pcgf3/5 interactome and identified several novel Pcgf3/5 interactors. These included testis-expressed 10 (Tex10), which may directly contribute to transcriptional activation via the transcriptional co-activator p300. Furthermore, Pcgf3/5 deletion in ES cells substantially reduced the occupancy of Tex10 and p300 at target genes. Finally, we demonstrated that Pcgf3/5 are essential for regulating global levels of the histone modifier H2AK119ub1 in ES cells. Our findings establish Pcgf3/5 as transcriptional activators that interact with Tex10 and p300 in ES cells and point to redundant activity of Pcgf3/5 in pluripotency maintenance.

show abstract

“…At molecular level, Pcgf6 null ES cells showed overexpression of genes involved in differentiation, especially in spermatogenesis and tend to differentiate towards endoderm when they formed EBs in vitro. Importantly, Pcgf 6 deletion in MEF did not have any effect on proliferation, suggesting that PCGF6 is important for stem cell-renewal but its function is dispensable in differentiated cells [339]. The genomic binding profile of PCGF6 is similar to TRXG activators and coincides to active chromatin marks.…”

Section: Ncprc13 and Ncprc15mentioning

confidence: 82%

“…CpG targeted ncPRC1.1 complexes are also active during pre-implantation development and play important role in establishing cellular plasticity of the early embryo [351]. Different ncPRC complexes, such as ncPRC1.6, can use independent targeting mechanism, and have a different, but just as important role in ES cell fate regulation [339]. The ncPRC1.6 complex does not harbor KDM2B or FBX10 subunits.…”

Section: Targeting Of Different Ncprc1 Complexesmentioning

confidence: 99%

From Flies to Mice: The Emerging Role of Non-Canonical PRC1 Members in Mammalian Development

2018

View full text Add to dashboard Cite

Originally two types of Polycomb Repressive Complexes (PRCs) were described, canonical PRC1 (cPRC1) and PRC2. Recently, a versatile set of complexes were identified and brought up several dilemmas in PRC mediated repression. These new class of complexes were named as non-canonical PRC1s (ncPRC1s). Both cPRC1s and ncPRC1s contain Ring finger protein (RING1, RNF2) and Polycomb group ring finger catalytic (PCGF) core, but in ncPRCs, RING and YY1 binding protein (RYBP), or YY1 associated factor 2 (YAF2), replaces the Chromobox (CBX) and Polyhomeotic (PHC) subunits found in cPRC1s. Additionally, ncPRC1 subunits can associate with versatile accessory proteins, which determine their functional specificity. Homozygous null mutations of the ncPRC members in mice are often lethal or cause infertility, which underlines their essential functions in mammalian development. In this review, we summarize the mouse knockout phenotypes of subunits of the six major ncPRCs. We highlight several aspects of their discovery from fly to mice and emerging role in target recognition, embryogenesis and cell-fate decision making. We gathered data from stem cell mediated in vitro differentiation assays and genetically engineered mouse models. Accumulating evidence suggests that ncPRC1s play profound role in mammalian embryogenesis by regulating gene expression during lineage specification of pluripotent stem cells.

show abstract

Essential Role for Polycomb Group Protein Pcgf6 in Embryonic Stem Cell Maintenance and a Noncanonical Polycomb Repressive Complex 1 (PRC1) Integrity

Cited by 50 publications

References 53 publications

Loss of Polycomb Group Protein Pcgf1 Severely Compromises Proper Differentiation of Embryonic Stem Cells

Loss of Polycomb Group Protein Pcgf1 Severely Compromises Proper Differentiation of Embryonic Stem Cells

Polycomb group RING finger proteins 3/5 activate transcription via an interaction with the pluripotency factor Tex10 in embryonic stem cells

From Flies to Mice: The Emerging Role of Non-Canonical PRC1 Members in Mammalian Development

Contact Info

Product

Resources

About