Silencing by small RNAs is linked to endosomal trafficking

Young Sik Lee; Sigal Pressman; Arlise P. Andress; Kevin Kim; Jamie L. White; Justin J. Cassidy; Xin Li; Kim Lubell; Do Hwan Lim; Ik Sang Cho; Kenji Nakahara; Jonathan B. Preall; Priya Bellare; Erik J. Sontheimer; Richard W. Carthew

doi:10.1038/ncb1930

Silencing by small RNAs is linked to endosomal trafficking

Young Sik Lee, Sigal Pressman, Arlise P. Andress, Kevin Kim, Jamie L. White, Justin J. Cassidy, Xin Li, Kim Lubell, Do Hwan Lim, Ik Sang Cho, Kenji Nakahara, Jonathan B. Preall, Priya Bellare, Erik J. Sontheimer, Richard W. Carthew

Division of Biotechnology

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

296 Citations (Scopus)

Abstract

Small RNAs direct RNA-induced silencing complexes (RISCs) to regulate stability and translation of mRNAs. RISCs associated with target mRNAs often accumulate in discrete cytoplasmic foci known as GW-bodies. However, RISC proteins can associate with membrane compartments such as the Golgi and endoplasmic reticulum. Here, we show that GW-bodies are associated with late endosomes (multivesicular bodies, MVBs). Blocking the maturation of MVBs into lysosomes by loss of the tethering factor HPS4 (ref. 5) enhances short interfering RNA (siRNA)- and micro RNA (miRNA)-mediated silencing in Drosophila melanogaster and humans. It also triggers over-accumulation of GW-bodies. Blocking MVB formation by ESCRT (endosomal sorting complex required for transport) depletion results in impaired miRNA silencing and loss of GW-bodies. These results indicate that active RISCs are physically and functionally coupled to MVBs. We further show that MVBs promote the competence of RISCs in loading small RNAs. We suggest that the recycling of RISCs is promoted by MVBs, resulting in RISCs more effectively engaging with small RNA effectors and possibly target RNAs. It may provide a means to enhance the dynamics of RNA silencing in the cytoplasm.

Original language	English
Pages (from-to)	1150-1156
Number of pages	7
Journal	Nature Cell Biology
Volume	11
Issue number	9
DOIs	https://doi.org/10.1038/ncb1930
Publication status	Published - 2009

Bibliographical note

Funding Information:
We thank R. Cagan, H. Folsch, P. Sharp, G. J. Hannon, Q. Liu, X. Wang, C. Horvath, G. Goshima, V. Helfand, M. Lowe, I. Hariharan, D. Bilder, J. Treisman, M. Siomi and J. Pham for reagents; H. Folsch, I. Fields, A. Komuro, and D. Harris for help with some of the experiments; H. Jiang for help with statistics and the Bloomington Stock Center and the Developmental Studies Hybridoma Bank for fly strains and antibodies. Y.S.L. was supported by a FRAXA postdoctoral Fellowship; J.J.C. was supported by the CMBD Training Grant and J.B.P. was supported by a Presidential Fellowship. This work was also supported by grants from the National Institutes of Health to E.J.S. (GM072830) and R.W.C. (GM77581, GM68743), and from a grant from the Korean BioGreen 21 Program to Y.S.L. (20070301034036).

ASJC Scopus subject areas

Cell Biology

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@article{24d1848810704d11a099e56a11fe8790,

title = "Silencing by small RNAs is linked to endosomal trafficking",

abstract = "Small RNAs direct RNA-induced silencing complexes (RISCs) to regulate stability and translation of mRNAs. RISCs associated with target mRNAs often accumulate in discrete cytoplasmic foci known as GW-bodies. However, RISC proteins can associate with membrane compartments such as the Golgi and endoplasmic reticulum. Here, we show that GW-bodies are associated with late endosomes (multivesicular bodies, MVBs). Blocking the maturation of MVBs into lysosomes by loss of the tethering factor HPS4 (ref. 5) enhances short interfering RNA (siRNA)- and micro RNA (miRNA)-mediated silencing in Drosophila melanogaster and humans. It also triggers over-accumulation of GW-bodies. Blocking MVB formation by ESCRT (endosomal sorting complex required for transport) depletion results in impaired miRNA silencing and loss of GW-bodies. These results indicate that active RISCs are physically and functionally coupled to MVBs. We further show that MVBs promote the competence of RISCs in loading small RNAs. We suggest that the recycling of RISCs is promoted by MVBs, resulting in RISCs more effectively engaging with small RNA effectors and possibly target RNAs. It may provide a means to enhance the dynamics of RNA silencing in the cytoplasm.",

author = "Lee, {Young Sik} and Sigal Pressman and Andress, {Arlise P.} and Kevin Kim and White, {Jamie L.} and Cassidy, {Justin J.} and Xin Li and Kim Lubell and Lim, {Do Hwan} and Cho, {Ik Sang} and Kenji Nakahara and Preall, {Jonathan B.} and Priya Bellare and Sontheimer, {Erik J.} and Carthew, {Richard W.}",

note = "Funding Information: We thank R. Cagan, H. Folsch, P. Sharp, G. J. Hannon, Q. Liu, X. Wang, C. Horvath, G. Goshima, V. Helfand, M. Lowe, I. Hariharan, D. Bilder, J. Treisman, M. Siomi and J. Pham for reagents; H. Folsch, I. Fields, A. Komuro, and D. Harris for help with some of the experiments; H. Jiang for help with statistics and the Bloomington Stock Center and the Developmental Studies Hybridoma Bank for fly strains and antibodies. Y.S.L. was supported by a FRAXA postdoctoral Fellowship; J.J.C. was supported by the CMBD Training Grant and J.B.P. was supported by a Presidential Fellowship. This work was also supported by grants from the National Institutes of Health to E.J.S. (GM072830) and R.W.C. (GM77581, GM68743), and from a grant from the Korean BioGreen 21 Program to Y.S.L. (20070301034036).",

year = "2009",

doi = "10.1038/ncb1930",

language = "English",

volume = "11",

pages = "1150--1156",

journal = "Nature Cell Biology",

issn = "1465-7392",

publisher = "Nature Publishing Group",

number = "9",

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T1 - Silencing by small RNAs is linked to endosomal trafficking

AU - Lee, Young Sik

AU - Pressman, Sigal

AU - Andress, Arlise P.

AU - Kim, Kevin

AU - White, Jamie L.

AU - Cassidy, Justin J.

AU - Li, Xin

AU - Lubell, Kim

AU - Lim, Do Hwan

AU - Cho, Ik Sang

AU - Nakahara, Kenji

AU - Preall, Jonathan B.

AU - Bellare, Priya

AU - Sontheimer, Erik J.

AU - Carthew, Richard W.

N1 - Funding Information: We thank R. Cagan, H. Folsch, P. Sharp, G. J. Hannon, Q. Liu, X. Wang, C. Horvath, G. Goshima, V. Helfand, M. Lowe, I. Hariharan, D. Bilder, J. Treisman, M. Siomi and J. Pham for reagents; H. Folsch, I. Fields, A. Komuro, and D. Harris for help with some of the experiments; H. Jiang for help with statistics and the Bloomington Stock Center and the Developmental Studies Hybridoma Bank for fly strains and antibodies. Y.S.L. was supported by a FRAXA postdoctoral Fellowship; J.J.C. was supported by the CMBD Training Grant and J.B.P. was supported by a Presidential Fellowship. This work was also supported by grants from the National Institutes of Health to E.J.S. (GM072830) and R.W.C. (GM77581, GM68743), and from a grant from the Korean BioGreen 21 Program to Y.S.L. (20070301034036).

PY - 2009

Y1 - 2009

N2 - Small RNAs direct RNA-induced silencing complexes (RISCs) to regulate stability and translation of mRNAs. RISCs associated with target mRNAs often accumulate in discrete cytoplasmic foci known as GW-bodies. However, RISC proteins can associate with membrane compartments such as the Golgi and endoplasmic reticulum. Here, we show that GW-bodies are associated with late endosomes (multivesicular bodies, MVBs). Blocking the maturation of MVBs into lysosomes by loss of the tethering factor HPS4 (ref. 5) enhances short interfering RNA (siRNA)- and micro RNA (miRNA)-mediated silencing in Drosophila melanogaster and humans. It also triggers over-accumulation of GW-bodies. Blocking MVB formation by ESCRT (endosomal sorting complex required for transport) depletion results in impaired miRNA silencing and loss of GW-bodies. These results indicate that active RISCs are physically and functionally coupled to MVBs. We further show that MVBs promote the competence of RISCs in loading small RNAs. We suggest that the recycling of RISCs is promoted by MVBs, resulting in RISCs more effectively engaging with small RNA effectors and possibly target RNAs. It may provide a means to enhance the dynamics of RNA silencing in the cytoplasm.

AB - Small RNAs direct RNA-induced silencing complexes (RISCs) to regulate stability and translation of mRNAs. RISCs associated with target mRNAs often accumulate in discrete cytoplasmic foci known as GW-bodies. However, RISC proteins can associate with membrane compartments such as the Golgi and endoplasmic reticulum. Here, we show that GW-bodies are associated with late endosomes (multivesicular bodies, MVBs). Blocking the maturation of MVBs into lysosomes by loss of the tethering factor HPS4 (ref. 5) enhances short interfering RNA (siRNA)- and micro RNA (miRNA)-mediated silencing in Drosophila melanogaster and humans. It also triggers over-accumulation of GW-bodies. Blocking MVB formation by ESCRT (endosomal sorting complex required for transport) depletion results in impaired miRNA silencing and loss of GW-bodies. These results indicate that active RISCs are physically and functionally coupled to MVBs. We further show that MVBs promote the competence of RISCs in loading small RNAs. We suggest that the recycling of RISCs is promoted by MVBs, resulting in RISCs more effectively engaging with small RNA effectors and possibly target RNAs. It may provide a means to enhance the dynamics of RNA silencing in the cytoplasm.

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U2 - 10.1038/ncb1930

DO - 10.1038/ncb1930

M3 - Article

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VL - 11

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JO - Nature Cell Biology

JF - Nature Cell Biology

IS - 9

ER -

Silencing by small RNAs is linked to endosomal trafficking

Abstract

Bibliographical note

ASJC Scopus subject areas

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