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The Isolation of Total and Membrane-Bound Polysomes from Arabidopsis and the Detection of Their Associated AGO1 and sRNAs

  • Shengben LiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1932)

Abstract

Polysomes play central roles in protein synthesis. Free polysomes are responsible for the translation of cytosolic and nuclear proteins, and the membrane-bound polysomes are responsible for the translation of transmembrane and secreted proteins. Accumulative evidence from recent studies shows that microRNAs are highly enriched on polysomes to regulate the expression of their own and target mRNAs. Here, we describe experimental protocols for the isolation of total and membrane-bound polysomes and the detection of the associated microRNAs and AGO1 proteins.

Key words

Total polysome Membrane-bound polysome microRNA AGO1 

References

  1. 1.
    Rogers K, Chen X (2013) Biogenesis, turnover, and mode of action of plant microRNAs. Plant Cell 25:2383–2399CrossRefGoogle Scholar
  2. 2.
    Li S, Le B, Ma X, You C, Yu Y, Zhang B, Liu L, Gao L, Shi T, Zhao Y, Mo B, Cao X, Chen X (2016) Biogenesis of phased siRNAs on membrane-bound polysomes in Arabidopsis. elife 5:e22750CrossRefGoogle Scholar
  3. 3.
    Stalder L, Heusermann W, Sokol L, Trojer D, Wirz J, Hean J, Fritzsche A, Aeschimann F, Pfanzagl V, Basselet P, Weiler J, Hintersteiner M, Morrissey DV, Meisner-Kober NC (2013) The rough endoplasmic reticulum is a central nucleation site of siRNA-mediated RNA silencing. EMBO J 32:1115–1127CrossRefGoogle Scholar
  4. 4.
    Wu PH, Isaji M, Carthew RW (2013) Functionally diverse microRNA effector complexes are regulated by extracellular signaling. Mol Cell 52:113–123CrossRefGoogle Scholar
  5. 5.
    Bazin J, Baerenfaller K, Gosai SJ, Gregory BD, Crespi M, Bailey-Serres J (2017) Global analysis of ribosome-associated noncoding RNAs unveils new modes of translational regulation. Proc Natl Acad Sci U S A 114:E10018–E10027CrossRefGoogle Scholar
  6. 6.
    Lauressergues D, Couzigou JM, Clemente HS, Martinez Y, Dunand C, Becard G, Combier JP (2015) Primary transcripts of microRNAs encode regulatory peptides. Nature 520:90–93CrossRefGoogle Scholar
  7. 7.
    Li S, Liu L, Zhuang X, Yu Y, Liu X, Cui X, Ji L, Pan Z, Cao X, Mo B, Zhang F, Raikhel N, Jiang L, Chen X (2013) MicroRNAs inhibit the translation of target mRNAs on the endoplasmic reticulum in Arabidopsis. Cell 153:562–574CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Nanjing Agricultural UniversityNanjingChina

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