Regulation of Motor Neuron Specification by Phosphorylation of Neurogenin 2

F.M. Kirby Neurobiology Center, Children's Hospital, and Departments of Neurology and Neurobiology, Harvard Medical School, 300 Longwood Avenue, Boston, MA 02115, USA.
Neuron (Impact Factor: 15.05). 05/2008; 58(1):65-77. DOI: 10.1016/j.neuron.2008.01.037
Source: PubMed


The mechanisms by which proneural basic helix-loop-helix (bHLH) factors control neurogenesis have been characterized, but it is not known how they specify neuronal cell-type identity. Here, we provide evidence that two conserved serine residues on the bHLH factor neurogenin 2 (Ngn2), S231 and S234, are phosphorylated during motor neuron differentiation. In knockin mice in which S231 and S234 of Ngn2 were mutated to alanines, neurogenesis occurs normally, but motor neuron specification is impaired. The phosphorylation of Ngn2 at S231 and S234 facilitates the interaction of Ngn2 with LIM homeodomain transcription factors to specify motor neuron identity. The phosphorylation-dependent cooperativity between Ngn2 and homeodomain transcription factors may be a general mechanism by which the activities of bHLH and homeodomain proteins are temporally and spatially integrated to generate the wide diversity of cell types that are a hallmark of the nervous system.

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    • "RESULTS Ascl1 is regulated by multisite phosphorylation Ascl1 is a core component of a variety of transcription factor protocols that have been used to drive transdifferentiation of mammalian fibroblasts directly into neurons (Vierbuchen et al., 2010; Caiazzo et al., 2011; Pang et al., 2011; Pfisterer et al., 2011; Yang et al., 2011; Torper et al., 2013), but the post-translational control of this protein is largely unknown. Phosphorylation of a number of conserved serineproline (SP) sites regulates the activity of Ngn2 (Neurog2 – Mouse Genome Informatics) and Olig2 basic helix-loop-helix (bHLH) proneural proteins (Ma et al., 2008; Ali et al., 2011; Gaber and Novitch, 2011). Ascl1 also contains multiple SP sites (supplementary material Fig. S1) that we hypothesized could be functionally modified through phosphorylation by proline-directed kinases. "
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    ABSTRACT: Generation of neurons from patient fibroblasts using a combination of developmentally defined transcription factors has great potential in disease modelling, as well as ultimately for use in regeneration and repair. However, generation of physiologically mature neurons in vitro remains problematic. Here we demonstrate the cell-cycle-dependent phosphorylation of a key reprogramming transcription factor, Ascl1, on multiple serine-proline sites. This multisite phosphorylation is a crucial regulator of the ability of Ascl1 to drive neuronal differentiation and maturation in vivo in the developing embryo; a phosphomutant form of Ascl1 shows substantially enhanced neuronal induction activity in Xenopus embryos. Mechanistically, we see that this un(der)phosphorylated Ascl1 is resistant to inhibition by both cyclin-dependent kinase activity and Notch signalling, both of which normally limit its neurogenic potential. Ascl1 is a central component of reprogramming transcription factor cocktails to generate neurons from human fibroblasts; the use of phosphomutant Ascl1 in place of the wild-type protein significantly promotes neuronal maturity after human fibroblast reprogramming in vitro. These results demonstrate that cell-cycle-dependent post-translational modification of proneural proteins directly regulates neuronal differentiation in vivo during development, and that this regulatory mechanism can be harnessed to promote maturation of neurons obtained by transdifferentiation of human cells in vitro.
    Development 05/2014; 141(11). DOI:10.1242/dev.106377 · 6.46 Impact Factor
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    • "Thus, further investigation may be warranted to determine whether a similar cooperative mechanism involving a yet to be identified Sox partner recruits Oct4 to genes that promote the mesodermal fate (Figure 1b). Finally, given that posttranslational modifications (e.g., phosphorylation) are also known to modulate the functional outcome of key transcription factors during neural tube development (Ma et al, 2008), it will be of interest to investigate whether such modifications can also affect the partnership of Oct4 with other lineage commitment factors. "
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    The EMBO Journal 03/2013; 32(7). DOI:10.1038/emboj.2013.48 · 10.43 Impact Factor
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    • "Interactions with partner proteins may modulate transcriptional activity of LIM-HD proteins (Matthews and Visvader, 2003). Furthermore, the post-translational modification of LIM-HD proteins or their interacting proteins may further influence their transcriptional activity or their subcellular locations (Ma et al., 2008; Petit et al., 2003). To achieve its diverse functions during cortical development, Lhx2 may regulate a variety of downstream genes, in part by interacting with different cofactors specific to each role for Lhx2 in corticogenesis. "
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