The Pluripotency Rheostat Nanog Functions as a Dimer

Adam Yates, Nicholas P. Mullin, Arthur J. Rowe, Bianca Nijmeijer, Douglas Colby, Paul N. Barlow, Malcolm D. Walkinshaw, Ian Chambers

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

The defining activity of the homeodomain protein Nanog is the ability to confer cytokine-independent self-renewal upon ES (embryonic stem) cells in which it is overexpressed. However, the biochemical basis by which Nanog achieves this function remains unknown. In the present study, we show that Nanog dimerizes through a functionally critical domain. Co-immunoprecipitation of Nanog molecules tagged with distinct epitopes demonstrates that Nanog self-associates through a region in which every fifth residue is tryptophan. In vitro binding experiments establish that this region participates directly in self-association. Moreover, analytical ultracentrifugation indicates that, in solution, Nanog is in equilibrium between monomeric and dimeric forms with a Kd of 3 μM. The functional importance of Nanog dimerization is established by ES cell colony-forming assays in which deletion of the tryptophan-repeat region eliminates the capacity of Nanog to direct LIF (leukaemia inhibitory factor)-independent self-renewal.

Original languageEnglish
Pages (from-to)227-231
Number of pages5
JournalBiochemical Journal
Volume411
Issue number2
DOIs
Publication statusPublished - 15 Apr 2008
Externally publishedYes

Fingerprint

Embryonic Stem Cells
Stem cells
Tryptophan
Dimers
Homeodomain Proteins
Leukemia Inhibitory Factor
Dimerization
Ultracentrifugation
Immunoprecipitation
Epitopes
Assays
Association reactions
Cytokines
Molecules
Experiments
In Vitro Techniques

Cite this

Yates, A., Mullin, N. P., Rowe, A. J., Nijmeijer, B., Colby, D., Barlow, P. N., ... Chambers, I. (2008). The Pluripotency Rheostat Nanog Functions as a Dimer. Biochemical Journal, 411(2), 227-231. https://doi.org/10.1042/BJ20080134
Yates, Adam ; Mullin, Nicholas P. ; Rowe, Arthur J. ; Nijmeijer, Bianca ; Colby, Douglas ; Barlow, Paul N. ; Walkinshaw, Malcolm D. ; Chambers, Ian. / The Pluripotency Rheostat Nanog Functions as a Dimer. In: Biochemical Journal. 2008 ; Vol. 411, No. 2. pp. 227-231.
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Yates, A, Mullin, NP, Rowe, AJ, Nijmeijer, B, Colby, D, Barlow, PN, Walkinshaw, MD & Chambers, I 2008, 'The Pluripotency Rheostat Nanog Functions as a Dimer', Biochemical Journal, vol. 411, no. 2, pp. 227-231. https://doi.org/10.1042/BJ20080134

The Pluripotency Rheostat Nanog Functions as a Dimer. / Yates, Adam; Mullin, Nicholas P.; Rowe, Arthur J.; Nijmeijer, Bianca; Colby, Douglas; Barlow, Paul N.; Walkinshaw, Malcolm D.; Chambers, Ian.

In: Biochemical Journal, Vol. 411, No. 2, 15.04.2008, p. 227-231.

Research output: Contribution to journalArticle

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Yates A, Mullin NP, Rowe AJ, Nijmeijer B, Colby D, Barlow PN et al. The Pluripotency Rheostat Nanog Functions as a Dimer. Biochemical Journal. 2008 Apr 15;411(2):227-231. https://doi.org/10.1042/BJ20080134