Sox2 interacts with Atoh1 and Huwe1 loci to regulate Atoh1 transcription and stability during hair cell differentiation
Data files
Jan 26, 2025 version files 20.94 KB
-
README.md
2.07 KB
-
Values_supporting_each_graph_PLOS_genetics.xlsx
18.86 KB
Abstract
Stem cell pluripotency gene Sox2 stimulates expression of proneural basic-helix-loop-helix transcription factor Atoh1. Sox2 is necessary for the development of cochlear hair cells and binds to the Atoh1 3’ enhancer to stimulate Atoh1 expression. We show here that Sox2 deletion in late embryogenesis results in the formation of extra hair cells, in contrast to the absence of hair cell development obtained after Sox2 knockout early in gestation. Sox2 overexpression decreased the level of Atoh1 protein despite an increase in Atoh1 mRNA. Sox2 upregulated E3 ubiquitin ligase, Huwe1, by direct binding to the Huwe1 gene. By upregulating its cognate E3 ligase, Sox2 disrupts the positive feedback loop through which Atoh1 protein increases the expression of Atoh1. We conclude that Sox2 initiates expression, while also limiting continued activity of bHLH transcription factor, Atoh1, and this inhibition represents a new mechanism for regulating the activity of this powerful initiator of hair cell development.
README: Sox2 Interacts with Atoh1 and Huwe1 Loci to Regulate Atoh1 Transcription and Stability During Hair Cell Differentiation
https://doi.org/10.5061/dryad.70rxwdc79
Description of the data and file structure
Regeneration of hair cells, the sensory cells of the cochlea, is a potential treatment for deafness. Here, we studied the regulation of Atoh1, a transcription factor required for the development of hair cells in the embryo. We discovered a dual role for Sox2 in determining the level of Atoh1 expression in the cochlea. Sox2 initiates the expression of Atoh1, while also imposing a limit on the level of Atoh1 protein through concurrent activation of the proteasome pathway. This mechanism of Atoh1 regulation explains the variable phenotypes induced by Sox2 deletion: a lack of hair cells when Sox2 was deleted early in embryogenesis was attributed to the missing stimulation by Sox2 of Atoh1 expression, while Sox2 deletion at later times in development resulted in an overproduction of hair cells due to decreased degradation of Atoh1 at the lowered level of E3 ubiquitin ligase Huwe1 ( HECT domain E3 ubiquitin ligase). This seemingly contradictory signaling enables Sox2 to direct the fate of inner ear progenitor cells through a brief burst of Atoh1 gene expression followed immediately by degradation of the protein. DOX- Doxycycline, MG132- Proteosome inhibitor.
Files and variables
File: Values_supporting_each_graph_PLOS_genetics.xlsx
Description: Numerical values of all points in figures
Variables
- Figure 1D: Cell number
- Figure 2A: Relative mRNA expression
- Figure 2C: Relative mRNA expression
- Figure 2F: Cell number
- Figure 2I: Cell number
- Figure 3B: Relative mRNA expression
- Figure 3H: Cell number
- Figure 3I: Cell number
- Figure 4B: Fold enrichment
- Figure 4D: Fold enrichment
- Figure 5A: Relative mRNA expression
- Figure 5E: Relative mRNA expression
- Figure 5F: Relative mRNA expression
- Figure 1S: Relative mRNA expression