RNA-directed DNA methylation involves co-transcriptional small-RNA-guided slicing of polymerase V transcripts in Arabidopsis

Liu, Wanlu1,2; Duttke, Sascha H.3,4,5,6; Hetzel, Jonathan3,4,5; Groth, Martin2; Feng, Suhua2,7; Gallego-Bartolome, Javier2; Zhong, Zhenhui2,8; Kuo, Hsuan Yu2; Wang, Zonghua8; Zhai, Jixian2,9; Chory, Joanne3,4,5; Jacobsen, Steven E.1,2,7,10

2018-03

10.1038/s41477-017-0100-y

Nature Plants   影响因子和分区

2055-026X

2055-0278

Small RNAs regulate chromatin modifications such as DNA methylation and gene silencing across eukaryotic genomes. In plants, RNA-directed DNA methylation (RdDM) requires 24-nucleotide small interfering RNAs (siRNAs) that bind to ARGONAUTE 4 (AGO4) and target genomic regions for silencing. RdDM also requires non-coding RNAs transcribed by RNA polymerase V (Pol V) that probably serve as scaffolds for binding of AGO4-siRNA complexes. Here, we used a modified global nuclear run-on protocol followed by deep sequencing to capture Pol V nascent transcripts genome-wide. We uncovered unique characteristics of Pol V RNAs, including a uracil (U) common at position 10. This uracil was complementary to the 5' adenine found in many AGO4-bound 24-nucleotide siRNAs and was eliminated in a siRNA-deficient mutant as well as in the ago4/6/9 triple mutant, suggesting that the +10 U signature is due to siRNA-mediated co-transcriptional slicing of Pol V transcripts. Expression of wild-type AGO4 in ago4/6/9 mutants was able to restore slicing of Pol V transcripts, but a catalytically inactive AGO4 mutant did not correct the slicing defect. We also found that Pol V transcript slicing required SUPPRESSOR OF TY INSERTION 5-LIKE (SPT5L), an elongation factor whose function is not well understood. These results highlight the importance of Pol V transcript slicing in RNA-mediated transcriptional gene silencing, which is a conserved process in many eukaryotes.

SCI

英语

其他

NIH[GM60398] ; NIH[R01GM094428] ; NIH[R01GM52413]

Plant Sciences

Plant Sciences

WOS:000428810500014

NATURE PUBLISHING GROUP

Web of Science

1.Univ Calif Los Angeles, Inst Mol Biol, Los Angeles, CA 90024 USA
2.Univ Calif Los Angeles, Dept Mol Cell & Dev Biol, Los Angeles, CA 90095 USA
3.Salk Inst Biol Studies, Plant Biol Lab, La Jolla, CA 92037 USA
4.Univ Calif San Diego, Div Biol Sci, La Jolla, CA 92093 USA
5.Salk Inst Biol Studies, Howard Hughes Med Inst, La Jolla, CA 92037 USA
6.Univ Calif San Diego, Sch Med, Dept Cellular & Mol Med, La Jolla, CA 92093 USA
7.Univ Calif Los Angeles, Eli & Edythe Broad Ctr Regenerat Med & Stem Cell, Los Angeles, CA 90095 USA
8.Fujian Agr & Forestry Univ, Coll Plant Protect, State Key Lab Ecol Pest Control Fujian & Taiwan C, Fuzhou, Fujian, Peoples R China
9.Southern Univ Sci & Technol, Dept Biol, Inst Plant & Food Sci, Shenzhen, Peoples R China
10.Univ Calif Los Angeles, Howard Hughes Med Inst, Los Angeles, CA 90024 USA

Liu, Wanlu,Duttke, Sascha H.,Hetzel, Jonathan,et al. RNA-directed DNA methylation involves co-transcriptional small-RNA-guided slicing of polymerase V transcripts in Arabidopsis[J]. Nature Plants,2018,4(3):181-188.

Liu, Wanlu.,Duttke, Sascha H..,Hetzel, Jonathan.,Groth, Martin.,Feng, Suhua.,...&Jacobsen, Steven E..(2018).RNA-directed DNA methylation involves co-transcriptional small-RNA-guided slicing of polymerase V transcripts in Arabidopsis.Nature Plants,4(3),181-188.

Liu, Wanlu,et al."RNA-directed DNA methylation involves co-transcriptional small-RNA-guided slicing of polymerase V transcripts in Arabidopsis".Nature Plants 4.3(2018):181-188.