ATP-dependent human RISC assembly pathways
Open accessAbstract
The assembly of RNA-induced silencing complex (RISC) is a key process in small RNA-mediated gene silencing. In humans, small interfering RNAs (siRNAs) and microRNAs (miRNAs) are incorporated into RISCs containing the Argonaute (AGO) subfamily proteins Ago1-4. Previous studies have proposed that, unlike Drosophila melanogaster RISC assembly pathways, human RISC assembly is coupled with dicing and is independent of ATP. Here we show by careful reexamination that, in humans, RISC assembly and dicing are uncoupled, and ATP greatly facilitates RISC loading of small-RNA duplexes. Moreover, all four human AGO proteins show remarkably similar structural preferences for small-RNA duplexes: central mismatches promote RISC loading, and seed or 3'-mid (guide position 12-15) mismatches facilitate unwinding. All these features of human AGO proteins are highly reminiscent of fly Ago1 but not fly Ago2.
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Both strands derived from the mature miRNA duplex can be loaded into the Argonaute family of proteins (AGO1-4 in humans) in an ATP-dependent manner.
“Both strands derived from the mature miRNA duplex can be loaded into the Argonaute (AGO) family of proteins (AGO1-4 in humans) in an ATP-dependent manner (32).”
From Overview of MicroRNA Biogenesis, Mechanisms of Actions, and Circulation (2018) · cites this paper as [32]
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- Authors
- Yoda, M., Kawamata, T., Paroo, Z., Ye, X., Iwasaki, S., Liu, Q., & Tomari, Y.
- Affiliations
- The University of Tokyo (JP)
- The University of Texas Southwestern Medical Center (US)
- Japan Science and Technology Agency (JP)
- Corresponding author
- Tomari, Yukihide
- Venue
- Nature Structural & Molecular Biology
- Published
- 2009-12-06
- Volume
- 17
- Issue
- 1
- Pages
- 17-23
- ISSN
- 1545-9985 (ISSN-L)
- 1545-9993
- PMC
- PMC2915567
- OpenAlex
- W1976048929
- Cited by
- 380
- References
- 48
- Citations by year
- 2026: 14 · 2025: 23 · 2024: 27 · 2023: 22 · 2022: 35 · 2021: 34 · 2020: 28 · 2019: 19 · 2018: 19 · 2017: 13 · 2016: 13 · 2015: 22 · 2014: 19 · 2013: 25 · 2012: 26
- FWCI
- 8.98
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Structural & Molecular Biology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Tokyo Tech Research Repository (Tokyo Institute of Technology) (submittedVersion)
- Funders
- National Institute of General Medical Sciences (R01GM078163)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Molecular Biology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Keywords
- RNA-induced silencing complex, Argonaute proteins, RNA interference, dicing, siRNA, microRNA, gene silencing
- MeSH terms
- Adenosine Triphosphate, HeLa Cells, Humans, Luciferases, Blotting, Western, Eukaryotic Initiation Factor-2, DNA Primers, RNA Interference, RNA, Small Interfering, RNA-Induced Silencing Complex, MicroRNAs, Eukaryotic Initiation Factors, Argonaute Proteins
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