Small RNAs (sRNAs) are loaded into ARGONAUTE (AGO) proteins to induce gene silencing. In plants, the 5′-terminal nucleotide is important for sRNA sorting into different AGOs. Here, we show that miRNA duplex structure also contributes to miRNA sorting. Base-pairing at the 15(th) nucleotide of a miRNA duplex is important for miRNA sorting in both Arabidopsis AGO1 and AGO2. AGO2 favors miRNA duplexes with no middle mismatches, whereas AGO1 tolerates, or prefers, duplexes with central mismatches. AGO structure modeling and mutational analyses reveal that the QF-V motif within the conserved PIWI domain contributes to recognition of base-pairing at the 15(th) nucleotide of a duplex, while the DDDE catalytic core of AtAGO2 is important for recognition of the central nucleotides. Finally, we rescued the adaxialized phenotype of ago1-12, which is largely due to miR165 loss-of-function, by changing miR165 duplex structure which we predict redirects it to AGO2

A Boland

A Carbonell

A Khvorova

A Mallory

A Pedretti

A Takeda

AC Mallory

AE Martínez de Alba

AL Eamens

B Czech

BR Brooks

C Notredame

C Poulsen

CA Kidner

CR Faehnle

DS Schwarz

E Elkayam

EJ Chapman

F Frank

FA Steiner

G Meister

H Vaucheret

HL Zhu

J-B Ma

JC Phillips

JJ Song

JR McConnell

K Arnold

K Förstemann

K Nakanishi

K Okamura

M Ghildiyal

N Maunoury

NT Schirle

PD Zamore

SB Li

SJ Mi

SL Ameres

T Kawamata

TA Montgomery

W Humphrey

XM Zhang

Y Endo

Y Tomari

Y Wang

PubMed

Xiaoming Zhang

DongDong Niu

Alberto Carbonell

Airong Wang

Angel Lee

Vinnary Tun

Zonghua Wang

James C. Carrington

Chia-en A. Chang

Hailing Jin

Crossref

Nature Communications

ARGONAUTE PIWI domain and microRNA duplex structure regulate small RNA sorting in Arabidopsis

[EN] Small RNAs (sRNAs) are loaded into ARGONAUTE (AGO) proteins to induce gene silencing. In plants, the 5¿-terminal nucleotide is important for sRNA sorting into different AGOs. Here we show that microRNA (miRNA) duplex structure also contributes to miRNA sorting. Base pairing at the 15th nucleotide of a miRNA duplex is important for miRNA sorting in both Arabidopsis ¿AGO1 and ¿AGO2. ¿AGO2 favours miRNA duplexes with no middle mismatches, whereas ¿AGO1 tolerates, or prefers, duplexes with central mismatches. AGO structure modelling and mutational analyses reveal that the QF-V motif within the conserved PIWI domain contributes to recognition of base pairing at the 15th nucleotide of a duplex, while the DDDE catalytic core of ¿AtAGO2 is important for recognition of the central nucleotides. Finally, we rescued the adaxialized phenotype of ¿ago1-12, which is largely due to miR165 loss-of-function, by changing miR165 duplex structure which we predict redirects it to ¿AGO2.We thank Yifan E. Lii, Patricia Springer and Hongwei Zhao for helpful comments; Yijun Qi, Xuemei Chen, David Baulcombe, Olivier Voinnet and Robert A. Martienssen for providing antibodies, constructs and seeds. This work was supported by an NIH (R01 GM093008), an NSF Career Award (MCB-0642843), an NSF Award (IOS-1257576) and an AES-CE Award (PPA-7517H) to H.J., and by NIH (AI043288) and NSF (MCB-0956526, MCB-1231726) grants to J.C.C. We thank Rizi Ai and Wanli You for constructing the protein and RNA complex structuresZhang, X.; Niu, D.; Carbonell, A.; Wang, A.; Lee, A.; Tun, V.; Wang, Z.... (2014). Argonaute PIWI domain and microRNA duplex structure contribute to small RNA sorting in Arabidopsis. Nature Communications. 5:1-11. https://doi.org/10.1038/ncomms64681115Zamore, P. D. & Haley, B. 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Zhang, Xiaoming

Niu, DongDong

CARBONELL, ALBERTO

Wang, Airong

Lee, Angel

Tun, Vinnary

Wang, Zonghua

Carrington, James C.

Chang, Chia-en A.

Jin, Hailing

RiuNet

Argonaute PIWI domain and microRNA duplex structure contribute to small RNA sorting in Arabidopsis.

Small RNAs (sRNAs) are loaded into ARGONAUTE (AGO) proteins to induce gene silencing. In plants, the 5'-terminal nucleotide is important for sRNA sorting into different AGOs. Here we show that microRNA (miRNA) duplex structure also contributes to miRNA sorting. Base pairing at the 15th nucleotide of a miRNA duplex is important for miRNA sorting in both Arabidopsis AGO1 and AGO2. AGO2 favours miRNA duplexes with no middle mismatches, whereas AGO1 tolerates, or prefers, duplexes with central mismatches. AGO structure modelling and mutational analyses reveal that the QF-V motif within the conserved PIWI domain contributes to recognition of base pairing at the 15th nucleotide of a duplex, while the DDDE catalytic core of AtAGO2 is important for recognition of the central nucleotides. Finally, we rescued the adaxialized phenotype of ago1-12, which is largely due to miR165 loss-of-function, by changing miR165 duplex structure which we predict redirects it to AGO2.Peer reviewe

Carbonell, Alberto

Carrington, James C

Chang, Chia-en A

Digital.CSIC

Small RNAs (sRNAs) are loaded into ARGONAUTE (AGO) proteins to induce gene silencing. In plants, the 5'-terminal nucleotide is important for sRNA sorting into different AGOs. Here we show that microRNA (miRNA) duplex structure also contributes to miRNA sorting. Base pairing at the 15th nucleotide of a miRNA duplex is important for miRNA sorting in both Arabidopsis AGO1 and AGO2. AGO2 favours miRNA duplexes with no middle mismatches, whereas AGO1 tolerates, or prefers, duplexes with central mismatches. AGO structure modelling and mutational analyses reveal that the QF-V motif within the conserved PIWI domain contributes to recognition of base pairing at the 15th nucleotide of a duplex, while the DDDE catalytic core of AtAGO2 is important for recognition of the central nucleotides. Finally, we rescued the adaxialized phenotype of ago1-12, which is largely due to miR165 loss-of-function, by changing miR165 duplex structure which we predict redirects it to AGO2

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ARGONAUTE PIWI domain and microRNA duplex structure regulate small RNA sorting in Arabidopsis

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