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The 3′ termini of mouse Piwi-interacting RNAs are 2′-O-methylated


Piwi-interacting RNAs (piRNAs) are a germline-specific class of small noncoding RNAs that are essential for spermatogenesis, but their function and biogenesis remain elusive. Here we report a post-transcriptional modification of mouse piRNAs. Mass spectrometric analysis reveals that the piRNAs tested are fully modified by 2′-O-methylation at their 3′ termini. This observation may provide a clue to the biogenesis and function of piRNAs in spermatogenesis.

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Figure 1: piRNAs have 3′-terminal nucleosides resistant to periodate oxidation.
Figure 2: LC/MS analysis reveals that 3′-termini of piRNAs are 2′-O-methylated.

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We thank Suzuki laboratory members for fruitful discussions and technical support, and Z. Mourelatos and Y. Kirino for communicating their experimental results to us before publication and for useful suggestions. This work was supported by a grant from the New Energy and Industrial Technology Development Organization (to Tsutomu Suzuki).

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Authors and Affiliations



T.O. prepared and analyzed piRNAs, assisted by K.M. Y.S. and Takeo Suzuki contributed to the mass spectrometric analysis and data interpretation. H.U. carried out computational analysis. Tsutomu Suzuki designed and supervised project and wrote the manuscript.

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Correspondence to Tsutomu Suzuki.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Total nucleotide analysis of the piRNA fraction. (PDF 126 kb)

Supplementary Fig. 2

LC/MS fragment analyses of the piRNA fraction digested by RNase A and RNase T1. (PDF 236 kb)

Supplementary Fig. 3

Mass spec de novo sequencing of 3'-terminal fragments from individual piRNAs digested by RNase A (a) or RNase T1 (b) (PDF 109 kb)

Supplementary Table 1

Quantification of nucleosides in piRNAs. (PDF 112 kb)

Supplementary Table 2

List of mouse piRNAs with each of the 3′-terminal fragments determined by mass spectrometric de novo sequencing. (PDF 122 kb)

Supplementary Methods and References (PDF 249 kb)

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Ohara, T., Sakaguchi, Y., Suzuki, T. et al. The 3′ termini of mouse Piwi-interacting RNAs are 2′-O-methylated. Nat Struct Mol Biol 14, 349–350 (2007).

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