Selective influence of Sox2 on POU transcription factor binding in embryonic and neural stem cells.
Embryonic stem cell (ESC) identity is orchestrated by co-operativity between the transcription factors (TFs) Sox2 and the class V POU-TF Oct4 at composite Sox/Oct motifs. Neural stem cells (NSCs) lack Oct4 but express Sox2 and class III POU-TFs Oct6, Brn1 and Brn2. This raises the question of how Sox2 interacts with POU-TFs to transcriptionally specify ESCs versus NSCs. Here, we show that Oct4 alone binds the Sox/Oct motif and the octamer-containing palindromic MORE equally well. Sox2 binding selectively increases the affinity of Oct4 for the Sox/Oct motif. In contrast, Oct6 binds preferentially to MORE and is unaffected by Sox2. ChIP-Seq in NSCs shows the MORE to be the most enriched motif for class III POU-TFs, including MORE subtypes, and that the Sox/Oct motif is not enriched. These results suggest that in NSCs, co-operativity between Sox2 and class III POU-TFs may not occur and that POU-TF-driven transcription uses predominantly the MORE cis architecture. Thus, distinct interactions between Sox2 and POU-TF subclasses distinguish pluripotent ESCs from multipotent NSCs, providing molecular insight into how Oct4 alone can convert NSCs to pluripotency.
Department of Chemistry, National University of Singapore, Singapore, Singapore Developmental Cellomics Laboratory, Genome Institute of Singapore, Singapore, Singapore MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, Edinburgh, UK.
Tapan Kumar
TK
Mistri
Developmental Cellomics Laboratory, Genome Institute of Singapore, Singapore, Singapore.
Arun George
AG
Devasia
Developmental Cellomics Laboratory, Genome Institute of Singapore, Singapore, Singapore.
Lee Thean
LT
Chu
Department of Chemistry, National University of Singapore, Singapore, Singapore.
Wei Ping
WP
Ng
MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, Edinburgh, UK.
Florian
F
Halbritter
MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, Edinburgh, UK.
Douglas
D
Colby
Division of Molecular Neurobiology, MRC-National Institute for Medical Research, Mill Hill, London, UK.
Ben
B
Martynoga
MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, Edinburgh, UK.
Simon R
SR
Tomlinson
MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, Edinburgh, UK ichambers@ed.ac.uk paul.robson@jax.org twohland@nus.edu.sg.
Ian
I
Chambers
Developmental Cellomics Laboratory, Genome Institute of Singapore, Singapore, Singapore Department of Biological Sciences, National University of Singapore, Singapore, Singapore The Jackson Laboratory for Genomic Medicine, Farmington, CT, USA ichambers@ed.ac.uk paul.robson@jax.org twohland@nus.edu.sg.
Paul
P
Robson
Department of Chemistry, National University of Singapore, Singapore, Singapore Department of Biological Sciences, National University of Singapore, Singapore, Singapore Centre for Bioimaging Sciences, National University of Singapore, Singapore, Singapore ichambers@ed.ac.uk paul.robson@jax.org twohland@nus.edu.sg.
Thorsten
T
Wohland
D000818:Animals; D004622:Embryo, Mammalian; D053595:Embryonic Stem Cells / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins / Q000235:genetics; D051379:Mice; D058953:Neural Stem Cells / Q000378:metabolism*; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D050809:POU Domain Factors / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D055748:SOXB1 Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic
D018398:Homeodomain Proteins; D050814:Octamer Transcription Factor-3; D050809:POU Domain Factors; D055748:SOXB1 Transcription Factors; D014157:Transcription Factors; C085076:transcription factor Brn-2
MORE; PORE; Sox/Oct motif; fluorescence correlation spectroscopy; fluorescent protein‐based electrophoretic mobility shift assay