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Mapping of INS promoter interactions reveals its role in long-range regulation of SYT8 transcription

Nature Structural & Molecular Biology volume 18, pages 372–378 (2011)Cite this article

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Abstract

Insulin (INS) synthesis and secretion from pancreatic β-cells are tightly regulated; their deregulation causes diabetes. Here we map INS-associated loci in human pancreatic islets by 4C and 3C techniques and show that the INS gene physically interacts with the SYT8 gene, located over 300 kb away. This interaction is elevated by glucose and accompanied by increases in SYT8 expression. Inactivation of the INS promoter by promoter-targeting siRNA reduces SYT8 gene expression. SYT8-INS interaction and SYT8 transcription are attenuated by CTCF depletion. Furthermore, SYT8 knockdown decreases insulin secretion in islets. These results reveal a nonredundant role for SYT8 in insulin secretion and indicate that the INS promoter acts from a distance to stimulate SYT8 transcription. This suggests a function for the INS promoter in coordinating insulin transcription and secretion through long-range regulation of SYT8 expression in human islets.

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Figure 1: 4C-Seq analysis reveals the association of SYT8 with INS gene in human pancreatic islets.
Figure 2: Glucose stimulates INS interactions with the SYT8-TNNI2 gene locus and increases SYT8 and TNNI2 gene expression in human islets.
Figure 3: The INS promoter positively regulates SYT8 and TNNI2 gene expression in human islets.
Figure 4: CTCF positively regulates SYT8 and TNNI2 gene expression in human islets and is important for the maintenance of the SYT8-INS interaction in human islets and human fibroblasts.
Figure 5: SYT8 is an important regulator of insulin secretion in human islets.

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Acknowledgements

We thank the Islet Cell Resource Centers Basic Science Islet Distribution Program for providing isolated human pancreatic islets. We thank K. Cui and D. Schones for assistance with the Solexa sequencing and pipeline analysis. This work was supported by National Institute of Diabetes and Digestive and Kidney Diseases and National Heart, Lung, and Blood Institute (US) Intramural Research Programs.

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Authors and Affiliations

  1. Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA

    Zhixiong Xu & Gary Felsenfeld

  2. Laboratory of Molecular Immunology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA

    Gang Wei, Iouri Chepelev & Keji Zhao

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  1. Zhixiong Xu
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  2. Gang Wei
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Contributions

Z.X. and G.F. designed the experiments; Z.X. conducted the experiments; G.W., I.C. and K.Z. conducted and analyzed Solexa DNA Sequencing experiments; Z.X. and G.F. wrote the manuscript.

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Correspondence to Gary Felsenfeld.

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Xu, Z., Wei, G., Chepelev, I. et al. Mapping of INS promoter interactions reveals its role in long-range regulation of SYT8 transcription. Nat Struct Mol Biol 18, 372–378 (2011). https://doi.org/10.1038/nsmb.1993

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