Optogenetic regulation of site-specific subtelomeric DNA-methylation

Samrat Roy Choudhury, Yi Cui, Anoop Narayanan, David P. Gilley, Nazmul Huda, Chiao Ling Lo, Feng C. Zhou, Dinesh Yernool, Joseph Irudayaraj

Research output: Contribution to journalArticle

Abstract

Telomere length homeostasis, critical for chromosomal integrity and genome stability, is controlled by intricate molecular regulatory machinery that includes epigenetic modifications. Here, we examine site-specific and spatiotemporal alteration of the subtelomeric methylation of CpG islands using optogenetic tools to understand the epigenetic regulatory mechanisms of telomere length maintenance. Human DNA methyltransferase3A (DNMT3A) were assembled selectively at chromosome ends by fusion to cryptochrome 2 protein (CRY2) and its interacting complement, the basic helix loop helix protein-1 (CIB1). CIB1 was fused to the telomere-associated protein telomere repeat binding factor-1 (TRF1), which localized the protein complex DNMT3A-CRY2 at telomeric regions upon excitation by blue-light monitored by singlemolecule fluorescence analyses. Increased methylation was achieved selectively at subtelomeric CpG sites on the six examined chromosome ends specifically after bluelight activation, which resulted in progressive increase in telomere length over three generations of HeLa cell replications. The modular design of the fusion constructs presented here allows for the selective substitution of other chromatin modifying enzymes and for loci-specific targeting to regulate the epigenetic pathways at telomeres and other selected genomic regions of interest.

Original languageEnglish (US)
Pages (from-to)50380-50391
Number of pages12
JournalOncotarget
Volume7
Issue number31
DOIs
StatePublished - Jan 1 2016
Externally publishedYes

Keywords

  • Optogenetics
  • Single-cells tools
  • Subtelomeric DNA-methylation
  • Telomere-elongation

ASJC Scopus subject areas

  • Oncology

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  • Cite this

    Choudhury, S. R., Cui, Y., Narayanan, A., Gilley, D. P., Huda, N., Lo, C. L., Zhou, F. C., Yernool, D., & Irudayaraj, J. (2016). Optogenetic regulation of site-specific subtelomeric DNA-methylation. Oncotarget, 7(31), 50380-50391. https://doi.org/10.18632/oncotarget.10394