Limiting Ago Protein Restricts RNAi and MicroRNA Biogenesis During Early Development in Xenopus Laevis

Overview

Journal Genes Dev

Specialty Molecular Biology

Date 2011 May 18

PMID 21576259

Citations 50

Authors

Elsebet Lund

Michael D Sheets

Susanne Blaser Imboden

James E Dahlberg

Affiliations

Soon will be listed here.

Abstract

We show that, in Xenopus laevis oocytes and early embryos, double-stranded exogenous siRNAs cannot function as microRNA (miRNA) mimics in either deadenylation or guided mRNA cleavage (RNAi). Instead, siRNAs saturate and inactivate maternal Argonaute (Ago) proteins, which are present in low amounts but are needed for Dicer processing of pre-miRNAs at the midblastula transition (MBT). Consequently, siRNAs impair accumulation of newly made miRNAs, such as the abundant embryonic pre-miR-427, but inhibition dissipates upon synthesis of zygotic Ago proteins after MBT. These effects of siRNAs, which are independent of sequence, result in morphological defects at later stages of development. The expression of any of several exogenous human Ago proteins, including catalytically inactive Ago2 (Ago2mut), can overcome the siRNA-mediated inhibition of miR-427 biogenesis and function. However, expression of wild-type, catalytically active hAgo2 is required to elicit RNAi in both early embryos and oocytes using either siRNA or endogenous miRNAs as guides. The lack of endogenous Ago2 endonuclease activity explains why these cells normally are unable to support RNAi. Expression of catalytically active exogenous Ago2, which appears not to perturb normal Xenopus embryonic development, can now be exploited for RNAi in this vertebrate model organism.

Citing Articles

CRISPR-RfxCas13d screening uncovers Bckdk as a post-translational regulator of the maternal-to-zygotic transition in teleosts.

Hernandez-Huertas L, Moreno-Sanchez I, Crespo-Cuadrado J, Vargas-Baco A, da Silva Pescador G, Santos-Pereira J bioRxiv. 2024; .

PMID: 38826327 PMC: 11142190. DOI: 10.1101/2024.05.22.595167.

Short 2'-O-methyl/LNA oligomers as highly-selective inhibitors of miRNA production in vitro and in vivo.

Koralewska N, Corradi E, Milewski M, Masante L, Szczepanska A, Kierzek R Nucleic Acids Res. 2024; 52(10):5804-5824.

PMID: 38676942 PMC: 11162791. DOI: 10.1093/nar/gkae284.

FAX-RIC enables robust profiling of dynamic RNP complex formation in multicellular organisms in vivo.

Na Y, Kim H, Choi Y, Shin S, Jung J, Kwon S Nucleic Acids Res. 2020; 49(5):e28.

PMID: 33332543 PMC: 7968992. DOI: 10.1093/nar/gkaa1194.

A CRISPR cut for messenger RNAs.

Leech R, Sampath K Lab Anim (NY). 2020; 49(11):317-319.

PMID: 33020605 DOI: 10.1038/s41684-020-00661-3.

The Evolution of Imprinted microRNAs and Their RNA Targets.

Haig D, Mainieri A Genes (Basel). 2020; 11(9).

PMID: 32899179 PMC: 7564603. DOI: 10.3390/genes11091038.

References

Rand T, Petersen S, Du F, Wang X . Argonaute2 cleaves the anti-guide strand of siRNA during RISC activation. Cell. 2005; 123(4):621-9. DOI: 10.1016/j.cell.2005.10.020. View

Audic Y, Anderson C, Bhatty R, Hartley R . Zygotic regulation of maternal cyclin A1 and B2 mRNAs. Mol Cell Biol. 2001; 21(5):1662-71. PMC: 86712. DOI: 10.1128/MCB.21.5.1662-1671.2001. View

Rudel S, Wang Y, Lenobel R, Korner R, Hsiao H, Urlaub H . Phosphorylation of human Argonaute proteins affects small RNA binding. Nucleic Acids Res. 2010; 39(6):2330-43. PMC: 3064767. DOI: 10.1093/nar/gkq1032. View

Gregory R, Chendrimada T, Cooch N, Shiekhattar R . Human RISC couples microRNA biogenesis and posttranscriptional gene silencing. Cell. 2005; 123(4):631-40. DOI: 10.1016/j.cell.2005.10.022. View

Matranga C, Tomari Y, Shin C, Bartel D, Zamore P . Passenger-strand cleavage facilitates assembly of siRNA into Ago2-containing RNAi enzyme complexes. Cell. 2005; 123(4):607-20. DOI: 10.1016/j.cell.2005.08.044. View

Kim V, Han J, Siomi M . Biogenesis of small RNAs in animals. Nat Rev Mol Cell Biol. 2009; 10(2):126-39. DOI: 10.1038/nrm2632. View

Eystathioy T, Chan E, Tenenbaum S, Keene J, Griffith K, Fritzler M . A phosphorylated cytoplasmic autoantigen, GW182, associates with a unique population of human mRNAs within novel cytoplasmic speckles. Mol Biol Cell. 2002; 13(4):1338-51. PMC: 102273. DOI: 10.1091/mbc.01-11-0544. View

Sheets M, Fox C, Hunt T, Vande Woude G, Wickens M . The 3'-untranslated regions of c-mos and cyclin mRNAs stimulate translation by regulating cytoplasmic polyadenylation. Genes Dev. 1994; 8(8):926-38. DOI: 10.1101/gad.8.8.926. View

Lykke-Andersen K, Gilchrist M, Grabarek J, Das P, Miska E, Zernicka-Goetz M . Maternal Argonaute 2 is essential for early mouse development at the maternal-zygotic transition. Mol Biol Cell. 2008; 19(10):4383-92. PMC: 2555945. DOI: 10.1091/mbc.e08-02-0219. View

10.

Diederichs S, Jung S, Rothenberg S, Smolen G, Mlody B, Haber D . Coexpression of Argonaute-2 enhances RNA interference toward perfect match binding sites. Proc Natl Acad Sci U S A. 2008; 105(27):9284-9. PMC: 2442125. DOI: 10.1073/pnas.0800803105. View

11.

Heasman J, Kofron M, Wylie C . Beta-catenin signaling activity dissected in the early Xenopus embryo: a novel antisense approach. Dev Biol. 2000; 222(1):124-34. DOI: 10.1006/dbio.2000.9720. View

12.

Wang L, Zhou J, Yao J, Lu D, Qiao X, Jia W . U6 promoter-driven siRNA injection has nonspecific effects in zebrafish. Biochem Biophys Res Commun. 2009; 391(3):1363-8. DOI: 10.1016/j.bbrc.2009.12.065. View

13.

Maniataki E, Mourelatos Z . A human, ATP-independent, RISC assembly machine fueled by pre-miRNA. Genes Dev. 2005; 19(24):2979-90. PMC: 1315402. DOI: 10.1101/gad.1384005. View

14.

Wu L, Fan J, Belasco J . Importance of translation and nonnucleolytic ago proteins for on-target RNA interference. Curr Biol. 2008; 18(17):1327-32. PMC: 3690557. DOI: 10.1016/j.cub.2008.07.072. View

15.

Koller E, Propp S, Murray H, Lima W, Bhat B, Prakash T . Competition for RISC binding predicts in vitro potency of siRNA. Nucleic Acids Res. 2006; 34(16):4467-76. PMC: 1636362. DOI: 10.1093/nar/gkl589. View

16.

Piao X, Zhang X, Wu L, Belasco J . CCR4-NOT deadenylates mRNA associated with RNA-induced silencing complexes in human cells. Mol Cell Biol. 2010; 30(6):1486-94. PMC: 2832495. DOI: 10.1128/MCB.01481-09. View

17.

Giraldez A . microRNAs, the cell's Nepenthe: clearing the past during the maternal-to-zygotic transition and cellular reprogramming. Curr Opin Genet Dev. 2010; 20(4):369-75. PMC: 2908189. DOI: 10.1016/j.gde.2010.04.003. View

18.

Newport J, Kirschner M . A major developmental transition in early Xenopus embryos: I. characterization and timing of cellular changes at the midblastula stage. Cell. 1982; 30(3):675-86. DOI: 10.1016/0092-8674(82)90272-0. View

19.

Khan A, Betel D, Miller M, Sander C, Leslie C, Marks D . Transfection of small RNAs globally perturbs gene regulation by endogenous microRNAs. Nat Biotechnol. 2009; 27(6):549-55. PMC: 2782465. DOI: 10.1038/nbt.1543. View

20.

Cheloufi S, Dos Santos C, Chong M, Hannon G . A dicer-independent miRNA biogenesis pathway that requires Ago catalysis. Nature. 2010; 465(7298):584-9. PMC: 2995450. DOI: 10.1038/nature09092. View