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MBD6 is a direct target of Oct4 and controls the stemness and differentiation of adipose tissue-derived stem cells

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Abstract

Argonaute 2 (Ago2) is a pivotal regulator of cell fate in adult stem cells. Its expression is significantly downregulated in late passages of cells, concomitant with a prominent increase in Ago2 cytosolic localization in single cells. Nuclear localization of Ago2 is crucial for the survival, proliferation, and differentiation of hATSCs (human adipose tissue-derived stem cells), mediated by the specific binding of the regulatory regions of functional genes, which positively or negatively altered gene expression. Ago2 targets genes that control stemness, reactive oxygen species scavenging, and microRNA expression, all of which are crucial for hATSC survival and self-renewal. Ago2 promotes cell proliferation and self-renewal by activating the expression of octamer-binding transcription factor 4 (Oct4). We confirmed the direct regulation of Oct4 activity by Ago2, as indicated by the results of the ChIP analysis. Methyl-CpG-binding protein 6 (MBD6) was detected as an Oct4 regulatory gene. As predicted, knockdown of MBD6 expression attenuated cell proliferation and eventually induced cell death. We hypothesized that MBD6 functions downstream of Oct4 in the regulation of stemness-related genes, cell proliferation, self-renewal activity, and survival. MBD6 also promoted cell transdifferentiation into neural and endodermal β-cells while significantly attenuating differentiation into the mesodermal lineage. We demonstrate that MBD6 is regulated by Ago2 via an interaction with Oct4, which alters self-renewal and gene expression in hATSCs. MBD6 was promoted cell proliferation through a novel set of signal mediators that may influence differentiation by repressing MBD2 and MBD3, which are possibly recruited by germ cell nuclear factor (GCNF).

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Abbreviations

Ago2:

Argonaute2

BRCA2:

Breast cancer type 2 susceptibility protein

BrdU:

5-bromo-2′-deoxyuridine

CDK:

Cyclin-dependent kinases

CFU:

Colony forming unit

CM-Dil:

Chloromethyl-benzamidodialkylcarbocyanine

DCAF-DA:

1′, 7′-Dichlodihydrofluorescein diacetate

FOXG1:

Forkhead box protein G1

GFAP:

Glial fibrillary acidic protein

hATSC:

human Adipose Tissue-derived Stem Cell

JNK:

c-Jun N-terminal kinases

KLF4:

Krueppel-like factor 4

MAP2:

Microtubule-associated protein 2

MBD:

Methyl-CpG-binding protein

NF160:

Neurofilament 160

Oct4:

Octamer-binding transcription factor 4

PPARγ:

Peroxisome proliferator-activated receptor gamma

RISC:

RNA-induced silencing complex

ROS:

Reactive Oxygen Species

Runx3:

Runt-related transcription factor 3

SEPN1:

Selenoprotein N1

Sox2:

SRY (sex determing region Y)-box 2

STAT3:

Signal transducer and activator of transcription 3

Tuj:

beta III tubulin

USP44:

Upiquitin carboxyl-terminal hydrolase 44

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Acknowledgments

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea Government (MEST, 2010-0020265).

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Authors and Affiliations

  1. Laboratory of Stem Cell Biology, Department of Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Republic of Korea

    Jin Sun Jung, Min Ki Jee, Hyun Tae Cho, Jee In Choi, Young Bin Im, Oh Hyun Kwon & Soo Kyung Kang

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  1. Jin Sun Jung
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  2. Min Ki Jee
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Correspondence to Soo Kyung Kang.

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Jin Sun Jung and Min Ki Jee contributed equally to this work.

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Jung, J.S., Jee, M.K., Cho, H.T. et al. MBD6 is a direct target of Oct4 and controls the stemness and differentiation of adipose tissue-derived stem cells. Cell. Mol. Life Sci. 70, 711–728 (2013). https://doi.org/10.1007/s00018-012-1157-4

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