Abstract
Members of the GATA protein family play important roles in lineage specification and transdifferentiation. Previous reports show that some members of the GATA protein family can also induce pluripotency in somatic cells by substituting for Oct4, a key pluripotency-associated factor. However, the mechanism linking lineage-specifying cues and the activation of pluripotency remains elusive. Here, we report that all GATA family members can substitute for Oct4 to induce pluripotency. We found that all members of the GATA family could inhibit the overrepresented ectodermal-lineage genes, which is consistent with previous reports indicating that a balance of different lineage-specifying forces is important for the restoration of pluripotency. A conserved zinc-finger DNA-binding domain in the C-terminus is critical for the GATA family to induce pluripotency. Using RNA-seq and ChIP-seq, we determined that the pluripotency-related gene Sall4 is a direct target of GATA family members during reprogramming and serves as a bridge linking the lineage-specifying GATA family to the pluripotency circuit. Thus, the GATA family is the first protein family of which all members can function as inducers of the reprogramming process and can substitute for Oct4. Our results suggest that the role of GATA family in reprogramming has been underestimated and that the GATA family may serve as an important mediator of cell fate conversion.
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Acknowledgements
We thank Yang Zhao and Jun Xu for helpful discussions. This work was supported by the National Basic Research Program of China (973 Program; 2012CB966401), the National Natural Science Foundation of China (91319305), the National Science and Technology Major Project (2013ZX10001003), the Ministry of Science and Technology of China (2013DFG30680) and the Ministry of Education of China (111 Project) to HD, and the Strategic Priority Research Program of the Chinese Academy of Sciences (XDA01040407), the National Natural Science Foundation of China (91019024 and 31100558) and the Hundred Talents Program of the Chinese Academy of Sciences to YS.
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Supplementary information
Supplementary information, Figure S1
Characterization of GATA-reprogrammed iPSCs. (PDF 6915 kb)
Supplementary information, Figure S2
Quantitative RT-PCR analysis of the expression of endogenous ECT related genes relative to the expression of SKM. (PDF 933 kb)
Supplementary information, Figure S3
A schematic diagram to illustrate zinc-finger domains in GATA family members. (PDF 386 kb)
Supplementary information, Figure S4
Strategies for mutation of GATA family members. (PDF 8061 kb)
Supplementary information, Figure S5
Quantitative RT-PCR analysis of the expression of endogenous Dlx3 and Lhx5 genes relative to the expression in SKM, G1D, G2D, G3D, G4D, G5D, and G6D represent corresponding GATA-Double mutations. (PDF 1166 kb)
Supplementary information, Figure S6
Venn diagram illustrating the overlap genes during secondary MEFs reprogramming. (PDF 1606 kb)
Supplementary information, Figure S7
Sall4 substitutes for Oct4 in MEF reprogramming to pluripotency. (PDF 3078 kb)
Supplementary information, Figure S8
ChIP-seq binding profiles for Sox2 and Nanog loci using secondary MEFs. (PDF 1800 kb)
Supplementary information, Figure S9
Western blot analysis of SALL4 knockdown in different GATA family member-mediated secondary MEF reprogramming. (PDF 1332 kb)
Supplementary information, Table S1
Number of reads (total reads, mapped reads, unique reads) in RNA-seq (GATA4) (PDF 4719 kb)
Supplementary information, Table S2
Distribution of all expressed genes in different expression levels (GATA4) (PDF 2268 kb)
Supplementary information, Table S3
Number of reads (total reads, mapped reads, unique reads) in RNA-seq (GATA6) (PDF 5183 kb)
Supplementary information, Table S4
Distribution of all expressed genes in different expression levels (GATA6) (PDF 2451 kb)
Supplementary information, Table S5
Number of reads (total reads, mapped reads, unique reads) in ChIP-seq (PDF 4465 kb)
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Shu, J., Zhang, K., Zhang, M. et al. GATA family members as inducers for cellular reprogramming to pluripotency. Cell Res 25, 169–180 (2015). https://doi.org/10.1038/cr.2015.6
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DOI: https://doi.org/10.1038/cr.2015.6
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