Roles for Drosophila cap1 2′-O-ribose methyltransferase in the small RNA silencing pathway associated with Argonaute 2

Seungjae Lee; Jae Sang Hong; Do Hwan Lim; Young Sik Lee

doi:10.1016/j.ibmb.2020.103415

Roles for Drosophila cap1 2′-O-ribose methyltransferase in the small RNA silencing pathway associated with Argonaute 2

Seungjae Lee, Jae Sang Hong, Do Hwan Lim, Young Sik Lee

Research output: Contribution to journal › Article › peer-review

4 Citations (Scopus)

Abstract

Cap1 2′-O-ribose methyltransferase (CMTR1) modifies RNA transcripts containing the 7-methylguanosine cap via 2′-O-ribose methylation of the first transcribed nucleotide, yielding cap1 structures. However, the role of CMTR1 in small RNA-mediated gene silencing remains unknown. Here, we identified and characterized a Drosophila CMTR1 gene (dCMTR1) mutation. We found that the catalytic activity of dCMTR1 was involved in the biogenesis of small interfering RNAs (siRNAs) but not microRNAs. Additionally, dCMTR1 interacted with R2D2, a key component for the assembly of the RNA-induced silencing complex (RISC) containing Argonaute 2 (Ago2). Consistent with this finding, loss of dCMTR1 function impaired RISC assembly by inhibiting the unwinding of Ago2-bound siRNA duplexes, thus preventing the retention of the guide strand. Moreover, dCMTR1 is unlikely to modify siRNAs during RISC assembly. Collectively, our data indicate that dCMTR1 is a positive regulator of the small RNA pathway associated with Ago2 with roles in both siRNA biogenesis and RISC assembly.

Original language	English
Article number	103415
Journal	Insect Biochemistry and Molecular Biology
Volume	123
DOIs	https://doi.org/10.1016/j.ibmb.2020.103415
Publication status	Published - 2020 Aug

Bibliographical note

Funding Information:
We thank the Bloomington Drosophila Stock Center for providing us with fly strains. We also thank M. Siomi for providing antibodies, a fly strain, and a construct expressing GST-Pimet, and G. Hannon for providing antibodies. This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education ( NRF-2018R1D1A1B07047792 to Y.S.L.); Korea University Grants (to Y.S.L. and D.-H.L.); and Korea University Future Research Grants (to Y.S.L.).

Publisher Copyright:
© 2020 Elsevier Ltd

Keywords

Ago2
CMTR1
Drosophila
RISC
cap1
siRNA

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Insect Science

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10.1016/j.ibmb.2020.103415

Cite this

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title = "Roles for Drosophila cap1 2′-O-ribose methyltransferase in the small RNA silencing pathway associated with Argonaute 2",

abstract = "Cap1 2′-O-ribose methyltransferase (CMTR1) modifies RNA transcripts containing the 7-methylguanosine cap via 2′-O-ribose methylation of the first transcribed nucleotide, yielding cap1 structures. However, the role of CMTR1 in small RNA-mediated gene silencing remains unknown. Here, we identified and characterized a Drosophila CMTR1 gene (dCMTR1) mutation. We found that the catalytic activity of dCMTR1 was involved in the biogenesis of small interfering RNAs (siRNAs) but not microRNAs. Additionally, dCMTR1 interacted with R2D2, a key component for the assembly of the RNA-induced silencing complex (RISC) containing Argonaute 2 (Ago2). Consistent with this finding, loss of dCMTR1 function impaired RISC assembly by inhibiting the unwinding of Ago2-bound siRNA duplexes, thus preventing the retention of the guide strand. Moreover, dCMTR1 is unlikely to modify siRNAs during RISC assembly. Collectively, our data indicate that dCMTR1 is a positive regulator of the small RNA pathway associated with Ago2 with roles in both siRNA biogenesis and RISC assembly.",

keywords = "Ago2, CMTR1, Drosophila, RISC, cap1, siRNA",

author = "Seungjae Lee and Hong, {Jae Sang} and Lim, {Do Hwan} and Lee, {Young Sik}",

note = "Funding Information: We thank the Bloomington Drosophila Stock Center for providing us with fly strains. We also thank M. Siomi for providing antibodies, a fly strain, and a construct expressing GST-Pimet, and G. Hannon for providing antibodies. This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education ( NRF-2018R1D1A1B07047792 to Y.S.L.); Korea University Grants (to Y.S.L. and D.-H.L.); and Korea University Future Research Grants (to Y.S.L.). Publisher Copyright: {\textcopyright} 2020 Elsevier Ltd",

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AU - Lee, Young Sik

N1 - Funding Information: We thank the Bloomington Drosophila Stock Center for providing us with fly strains. We also thank M. Siomi for providing antibodies, a fly strain, and a construct expressing GST-Pimet, and G. Hannon for providing antibodies. This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education ( NRF-2018R1D1A1B07047792 to Y.S.L.); Korea University Grants (to Y.S.L. and D.-H.L.); and Korea University Future Research Grants (to Y.S.L.). Publisher Copyright: © 2020 Elsevier Ltd

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N2 - Cap1 2′-O-ribose methyltransferase (CMTR1) modifies RNA transcripts containing the 7-methylguanosine cap via 2′-O-ribose methylation of the first transcribed nucleotide, yielding cap1 structures. However, the role of CMTR1 in small RNA-mediated gene silencing remains unknown. Here, we identified and characterized a Drosophila CMTR1 gene (dCMTR1) mutation. We found that the catalytic activity of dCMTR1 was involved in the biogenesis of small interfering RNAs (siRNAs) but not microRNAs. Additionally, dCMTR1 interacted with R2D2, a key component for the assembly of the RNA-induced silencing complex (RISC) containing Argonaute 2 (Ago2). Consistent with this finding, loss of dCMTR1 function impaired RISC assembly by inhibiting the unwinding of Ago2-bound siRNA duplexes, thus preventing the retention of the guide strand. Moreover, dCMTR1 is unlikely to modify siRNAs during RISC assembly. Collectively, our data indicate that dCMTR1 is a positive regulator of the small RNA pathway associated with Ago2 with roles in both siRNA biogenesis and RISC assembly.

AB - Cap1 2′-O-ribose methyltransferase (CMTR1) modifies RNA transcripts containing the 7-methylguanosine cap via 2′-O-ribose methylation of the first transcribed nucleotide, yielding cap1 structures. However, the role of CMTR1 in small RNA-mediated gene silencing remains unknown. Here, we identified and characterized a Drosophila CMTR1 gene (dCMTR1) mutation. We found that the catalytic activity of dCMTR1 was involved in the biogenesis of small interfering RNAs (siRNAs) but not microRNAs. Additionally, dCMTR1 interacted with R2D2, a key component for the assembly of the RNA-induced silencing complex (RISC) containing Argonaute 2 (Ago2). Consistent with this finding, loss of dCMTR1 function impaired RISC assembly by inhibiting the unwinding of Ago2-bound siRNA duplexes, thus preventing the retention of the guide strand. Moreover, dCMTR1 is unlikely to modify siRNAs during RISC assembly. Collectively, our data indicate that dCMTR1 is a positive regulator of the small RNA pathway associated with Ago2 with roles in both siRNA biogenesis and RISC assembly.

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Roles for Drosophila cap1 2′-O-ribose methyltransferase in the small RNA silencing pathway associated with Argonaute 2

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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