Prolyl hydroxylase 3 controls the intestine goblet cell generation through stabilizing ATOH1

Abstract

Intestinal epithelia self-renew constantly and generate differentiated cells such as secretary goblet cells. The intestine goblet cells secrete gel-forming mucins that form mucus to create a barrier of defense. We reported previously that loss of prolyl hydroxylase (PHD) 3 led to disruption of the intestinal epithelial barrier function. However, the underlying mechanism remains elusive. Here, we demonstrate that PHD3 controls the generation of intestine goblet cell. We found that genetic ablation of Phd3 in mice intestine epithelial cells reduced the amount of goblet cells. Mechanistically, PHD3 bounds the E3 ubiquitin ligase HUWE1 and prevented HUWE1 from mediating ubiquitination and degradation of ATOH1, an essential driver for goblet cell differentiation. The prolyl hydroxylase activity-deficient variant PHD3(H196A) also prevented ATOH1 destruction. A genetic intestine epithelial PHD3(H196A)-knockin had no effect on ATOH1 expression or goblet cell amount in mice, suggesting that the PHD3 prolyl hydroxylase activity is dispensable for its ability to control ATOH1 expression and goblet cell generation. In dextran sulfate sodium (DSS)-induced experimental colitis, PHD3-knockout rather than PHD3(H196A)-knockin sensitized the mice to DSS treatment. Our results reveal an additional critical mechanism underlying the regulation of ATOH1 expression and goblet cell generation and highlight that PHD3 plays a role in controlling intestine goblet cell generation in a hydroxylase-independent manner.

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Fig. 1: PHD3 deficiency reduced goblet cell number in mice intestines.
Fig. 2: PHD3 stabilized ATOH1.
Fig. 3: PHD3 bound HUWE1.
Fig. 4: PHD3 blocked the HUWE1–ATOH1 interaction.
Fig. 5: PHD3 controlled goblet cell generation independent of its hydroxylase activity.
Fig. 6: PHD(H196A) knock-in had little effect on DSS-induced colitis.

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Acknowledgements

This work was supported by China Ministry of Science and Technology (2017YFD0400206), National Natural Science Foundation of China (31470769, 31670785), and Shanghai Ministry of Science and Technology (16JC1406100).

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Correspondence to Jing Fang.

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Xu, Y., Gao, Q., Zhang, J. et al. Prolyl hydroxylase 3 controls the intestine goblet cell generation through stabilizing ATOH1. Cell Death Differ (2020). https://doi.org/10.1038/s41418-020-0490-7

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