The Role of Dicer Protein Partners in the Processing of MicroRNA Precursors

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2011 Dec 14

PMID 22163034

Citations 64

Authors

Edyta Koscianska

Julia Starega-Roslan

Wlodzimierz J Krzyzosiak

Affiliations

Soon will be listed here.

Abstract

One of the cellular functions of the ribonuclease Dicer is to process microRNA precursors (pre-miRNAs) into mature microRNAs (miRNAs). Human Dicer performs this function in cooperation with its protein partners, AGO2, PACT and TRBP. The exact role of these accessory proteins in Dicer activity is still poorly understood. In this study, we used the northern blotting technique to investigate pre-miRNA cleavage efficiency and specificity after depletion of AGO2, PACT and TRBP by RNAi. The results showed that the inhibition of either Dicer protein partner substantially affected not only miRNA levels but also pre-miRNA levels, and it had a rather minor effect on the specificity of Dicer cleavage. The analysis of the Dicer cleavage products generated in vitro revealed the presence of a cleavage intermediate when pre-miRNA was processed by recombinant Dicer alone. This intermediate was not observed during pre-miRNA cleavage by endogenous Dicer. We demonstrate that AGO2, PACT and TRBP were required for the efficient functioning of Dicer in cells, and we suggest that one of the roles of these proteins is to assure better synchronization of cleavages triggered by two RNase III domains of Dicer.

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References

Koscianska E, Starega-Roslan J, Czubala K, Krzyzosiak W . High-resolution northern blot for a reliable analysis of microRNAs and their precursors. ScientificWorldJournal. 2011; 11:102-17. PMC: 5720006. DOI: 10.1100/tsw.2011.11. View

Ruby J, Jan C, Player C, Axtell M, Lee W, Nusbaum C . Large-scale sequencing reveals 21U-RNAs and additional microRNAs and endogenous siRNAs in C. elegans. Cell. 2006; 127(6):1193-207. DOI: 10.1016/j.cell.2006.10.040. View

Nelson P, de Planell-Saguer M, Lamprinaki S, Kiriakidou M, Zhang P, ODoherty U . A novel monoclonal antibody against human Argonaute proteins reveals unexpected characteristics of miRNAs in human blood cells. RNA. 2007; 13(10):1787-92. PMC: 1986805. DOI: 10.1261/rna.646007. View

Diederichs S, Haber D . Dual role for argonautes in microRNA processing and posttranscriptional regulation of microRNA expression. Cell. 2007; 131(6):1097-108. DOI: 10.1016/j.cell.2007.10.032. 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

Landgraf P, Rusu M, Sheridan R, Sewer A, Iovino N, Aravin A . A mammalian microRNA expression atlas based on small RNA library sequencing. Cell. 2007; 129(7):1401-14. PMC: 2681231. DOI: 10.1016/j.cell.2007.04.040. View

OCarroll D, Mecklenbrauker I, Das P, Santana A, Koenig U, Enright A . A Slicer-independent role for Argonaute 2 in hematopoiesis and the microRNA pathway. Genes Dev. 2007; 21(16):1999-2004. PMC: 1948855. DOI: 10.1101/gad.1565607. View

Zhang H, Kolb F, Jaskiewicz L, Westhof E, Filipowicz W . Single processing center models for human Dicer and bacterial RNase III. Cell. 2004; 118(1):57-68. DOI: 10.1016/j.cell.2004.06.017. View

Leuschner P, Martinez J . In vitro analysis of microRNA processing using recombinant Dicer and cytoplasmic extracts of HeLa cells. Methods. 2007; 43(2):105-9. DOI: 10.1016/j.ymeth.2007.04.005. View

10.

Kok K, Ng M, Ching Y, Jin D . Human TRBP and PACT directly interact with each other and associate with dicer to facilitate the production of small interfering RNA. J Biol Chem. 2007; 282(24):17649-57. DOI: 10.1074/jbc.M611768200. View

11.

Starega-Roslan J, Koscianska E, Kozlowski P, Krzyzosiak W . The role of the precursor structure in the biogenesis of microRNA. Cell Mol Life Sci. 2011; 68(17):2859-71. PMC: 3155042. DOI: 10.1007/s00018-011-0726-2. View

12.

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

13.

Landthaler M, Yalcin A, Tuschl T . The human DiGeorge syndrome critical region gene 8 and Its D. melanogaster homolog are required for miRNA biogenesis. Curr Biol. 2004; 14(23):2162-7. DOI: 10.1016/j.cub.2004.11.001. View

14.

Park J, Heo I, Tian Y, Simanshu D, Chang H, Jee D . Dicer recognizes the 5' end of RNA for efficient and accurate processing. Nature. 2011; 475(7355):201-5. PMC: 4693635. DOI: 10.1038/nature10198. View

15.

Yi R, OCarroll D, Pasolli H, Zhang Z, Dietrich F, Tarakhovsky A . Morphogenesis in skin is governed by discrete sets of differentially expressed microRNAs. Nat Genet. 2006; 38(3):356-62. DOI: 10.1038/ng1744. View

16.

Starega-Roslan J, Krol J, Koscianska E, Kozlowski P, Szlachcic W, Sobczak K . Structural basis of microRNA length variety. Nucleic Acids Res. 2010; 39(1):257-68. PMC: 3017592. DOI: 10.1093/nar/gkq727. View

17.

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

18.

Lee Y, Hur I, Park S, Kim Y, Suh M, Kim V . The role of PACT in the RNA silencing pathway. EMBO J. 2006; 25(3):522-32. PMC: 1383527. DOI: 10.1038/sj.emboj.7600942. View

19.

Obernosterer G, Leuschner P, Alenius M, Martinez J . Post-transcriptional regulation of microRNA expression. RNA. 2006; 12(7):1161-7. PMC: 1484437. DOI: 10.1261/rna.2322506. View

20.

Shin Y, Bae E, Kim D . Inhibitory effect of ginsenoside Rg5 and its metabolite ginsenoside Rh3 in an oxazolone-induced mouse chronic dermatitis model. Arch Pharm Res. 2006; 29(8):685-90. DOI: 10.1007/BF02968253. View