open access

https://hdl.handle.net/2115/49934

DNA-RNA hybrid formation mediates RNAi-directed heterochromatin formation

英語

fission yeast

2012-03

Wiley-Blackwell

Genes to Cells

17(3)

218-233

Certain non-coding RNAs (ncRNAs) implicated in the regulation of chromatin structure associate with chromatin. During the formation of RNAi-directed heterochromatin in fission yeast, ncRNAs transcribed from heterochromatin are thought to recruit the RNAi machinery to chromatin for the formation of heterochromatin; however, the molecular details of this association are not clear. Here, using RNA-immunoprecipitation assay, we showed that the heterochromatic ncRNA associated with chromatin via the formation of a DNA:RNA hybrid and bound to the RNA-induced transcriptional silencing (RITS) complex. The presence of DNA:RNA hybrid in the cell was also confirmed by immunofluorescence analysis using anti-DNA:RNA hybrid antibody. Overexpression and depletion of RNase H in vivo decreased and increased the amount of DNA:RNA hybrid formed, respectively, and both disturbed heterochromatin. Moreover, DNA:RNA hybrid was formed on, and overexpression of RNase H inhibited the formation of, artificial heterochromatin induced by tethering of RITS to mRNA. These results indicate that heterochromatic ncRNAs are retained on chromatin via the formation of DNA:RNA hybrids and provide a platform for the RNAi-directed heterochromatin assembly and suggest that DNA:RNA hybrid formation plays a role in chromatic ncRNA function.

学術雑誌論文

Accepted Manuscript

The definitive version is available at www3.interscience.wiley.com

1356-9597