MIWI2 and MILI Have Differential Effects on piRNA Biogenesis and DNA Methylation
Abstract
In developing male germ cells, prospermatogonia, two Piwi proteins, MILI and MIWI2, use Piwi-interacting RNA (piRNA) guides to repress transposable element (TE) expression and ensure genome stability and proper gametogenesis. In addition to their roles in post-transcriptional TE repression, both proteins are required for DNA methylation of TE sequences. Here, we analyzed the effect of Miwi2 deficiency on piRNA biogenesis and transposon repression. Miwi2 deficiency had only a minor impact on piRNA biogenesis; however, the piRNA profile of Miwi2-knockout mice indicated overexpression of several LINE1 TE families that led to activation of the ping-pong piRNA cycle. Furthermore, we found that MILI and MIWI2 have distinct functions in TE repression in the nucleus. MILI is responsible for DNA methylation of a larger subset of TE families than MIWI2 is, suggesting that the proteins have independent roles in establishing DNA methylation patterns.
Additional Information
© 2015 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Received: May 14, 2015; revised: July 2, 2015; accepted: July 16, 2015; published: August 13, 2015. We thank Katalin Fejes Tóth and members of the Aravin lab for helpful discussion and comments on the manuscript. We thank Alyssa Maskell (Caltech) for assistance with mouse work. We thank Igor Antoshechkin (Caltech) for help with RNA-seq and Maria Ninova for help with genome annotation. This work was supported by grants from the NIH (R00 HD057233 and DP2 OD007371A) and by the Searle Scholar Award and Packard Fellowship (to A.A.A.). Author Contributions: S.A.M., D.P., and A.A.A. designed the experiments. D.P. performed the experiments, S.A.M. performed computational analysis of all deep-sequencing data, and G.K.M. performed ping-pong analysis. R.S. created bioinformatics tools for small RNA analysis. W.A.P. cloned and sequenced RNA-seq libraries from Mili mutant. S.A.M., D.P., and A.A.A. interpreted the data and wrote the manuscript.Attached Files
Published - 1-s2.0-S2211124715007962-main.pdf
Supplemental Material - mmc1.pdf
Supplemental Material - mmc2.pdf
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Additional details
- PMCID
- PMC4554733
- Eprint ID
- 59819
- Resolver ID
- CaltechAUTHORS:20150821-154811940
- R00 HD057233
- NIH
- DP2 OD007371A
- NIH
- Searle Scholars Program
- David and Lucile Packard Foundation
- Created
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2015-08-21Created from EPrint's datestamp field
- Updated
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2022-05-24Created from EPrint's last_modified field