The Hub Protein Loquacious Connects the MicroRNA and Short Interfering RNA Pathways in Mosquitoes

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2015 Mar 14

PMID 25765650

Citations 21

Authors

Mary Etna Haac

Michelle A E Anderson

Heather Eggleston

Kevin M Myles

Zach N Adelman

Affiliations

Soon will be listed here.

Abstract

Aedes aegypti mosquitoes vector several arboviruses of global health significance, including dengue viruses and chikungunya virus. RNA interference (RNAi) plays an important role in antiviral immunity, gene regulation and protection from transposable elements. Double-stranded RNA binding proteins (dsRBPs) are important for efficient RNAi; in Drosophila functional specialization of the miRNA, endo-siRNA and exo-siRNA pathway is aided by the dsRBPs Loquacious (Loqs-PB, Loqs-PD) and R2D2, respectively. However, this functional specialization has not been investigated in other dipterans. We were unable to detect Loqs-PD in Ae. aegypti; analysis of other dipteran genomes demonstrated that this isoform is not conserved outside of Drosophila. Overexpression experiments and small RNA sequencing following depletion of each dsRBP revealed that R2D2 and Loqs-PA cooperate non-redundantly in siRNA production, and that these proteins exhibit an inhibitory effect on miRNA levels. Conversely, Loqs-PB alone interacted with mosquito dicer-1 and was essential for full miRNA production. Mosquito Loqs interacted with both argonaute 1 and 2 in a manner independent of its interactions with dicer. We conclude that the functional specialization of Loqs-PD in Drosophila is a recently derived trait, and that in other dipterans, including the medically important mosquitoes, Loqs-PA participates in both the miRNA and endo-siRNA based pathways.

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References

Elbashir S, Lendeckel W, Tuschl T . RNA interference is mediated by 21- and 22-nucleotide RNAs. Genes Dev. 2001; 15(2):188-200. PMC: 312613. DOI: 10.1101/gad.862301. View

Sanchez-Vargas I, Travanty E, Keene K, Franz A, Beaty B, Blair C . RNA interference, arthropod-borne viruses, and mosquitoes. Virus Res. 2004; 102(1):65-74. DOI: 10.1016/j.virusres.2004.01.017. View

Gubler D . The changing epidemiology of yellow fever and dengue, 1900 to 2003: full circle?. Comp Immunol Microbiol Infect Dis. 2004; 27(5):319-30. DOI: 10.1016/j.cimid.2004.03.013. View

Hahn C, Hahn Y, Braciale T, Rice C . Infectious Sindbis virus transient expression vectors for studying antigen processing and presentation. Proc Natl Acad Sci U S A. 1992; 89(7):2679-83. PMC: 48725. DOI: 10.1073/pnas.89.7.2679. View

Tomari Y, Matranga C, Haley B, Martinez N, Zamore P . A protein sensor for siRNA asymmetry. Science. 2004; 306(5700):1377-80. DOI: 10.1126/science.1102755. View

Forstemann K, Tomari Y, Du T, Vagin V, Denli A, Bratu D . Normal microRNA maturation and germ-line stem cell maintenance requires Loquacious, a double-stranded RNA-binding domain protein. PLoS Biol. 2005; 3(7):e236. PMC: 1141267. DOI: 10.1371/journal.pbio.0030236. View

Jiang F, Ye X, Liu X, Fincher L, McKearin D, Liu Q . Dicer-1 and R3D1-L catalyze microRNA maturation in Drosophila. Genes Dev. 2005; 19(14):1674-9. PMC: 1176004. DOI: 10.1101/gad.1334005. View

Myles K, Kelly C, Ledermann J, Powers A . Effects of an opal termination codon preceding the nsP4 gene sequence in the O'Nyong-Nyong virus genome on Anopheles gambiae infectivity. J Virol. 2006; 80(10):4992-7. PMC: 1472075. DOI: 10.1128/JVI.80.10.4992-4997.2006. View

Liu X, Jiang F, Kalidas S, Smith D, Liu Q . Dicer-2 and R2D2 coordinately bind siRNA to promote assembly of the siRISC complexes. RNA. 2006; 12(8):1514-20. PMC: 1524895. DOI: 10.1261/rna.101606. View

10.

Ye X, Paroo Z, Liu Q . Functional anatomy of the Drosophila microRNA-generating enzyme. J Biol Chem. 2007; 282(39):28373-28378. DOI: 10.1074/jbc.M705208200. View

11.

Enserink M . Infectious diseases. Chikungunya: no longer a third world disease. Science. 2007; 318(5858):1860-1. DOI: 10.1126/science.318.5858.1860. View

12.

Czech B, Malone C, Zhou R, Stark A, Schlingeheyde C, Dus M . An endogenous small interfering RNA pathway in Drosophila. Nature. 2008; 453(7196):798-802. PMC: 2895258. DOI: 10.1038/nature07007. View

13.

Adelman Z, Anderson M, Morazzani E, Myles K . A transgenic sensor strain for monitoring the RNAi pathway in the yellow fever mosquito, Aedes aegypti. Insect Biochem Mol Biol. 2008; 38(7):705-13. PMC: 2518454. DOI: 10.1016/j.ibmb.2008.04.002. View

14.

Laraki G, Clerzius G, Daher A, Melendez-Pena C, Daniels S, Gatignol A . Interactions between the double-stranded RNA-binding proteins TRBP and PACT define the Medipal domain that mediates protein-protein interactions. RNA Biol. 2008; 5(2):92-103. DOI: 10.4161/rna.5.2.6069. View

15.

Myles K, Wiley M, Morazzani E, Adelman Z . Alphavirus-derived small RNAs modulate pathogenesis in disease vector mosquitoes. Proc Natl Acad Sci U S A. 2008; 105(50):19938-43. PMC: 2604946. DOI: 10.1073/pnas.0803408105. View

16.

Jinek M, Doudna J . A three-dimensional view of the molecular machinery of RNA interference. Nature. 2009; 457(7228):405-12. DOI: 10.1038/nature07755. View

17.

Sanchez-Vargas I, Scott J, Poole-Smith B, Franz A, Barbosa-Solomieu V, Wilusz J . Dengue virus type 2 infections of Aedes aegypti are modulated by the mosquito's RNA interference pathway. PLoS Pathog. 2009; 5(2):e1000299. PMC: 2633610. DOI: 10.1371/journal.ppat.1000299. View

18.

Langmead B, Trapnell C, Pop M, Salzberg S . Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol. 2009; 10(3):R25. PMC: 2690996. DOI: 10.1186/gb-2009-10-3-r25. View

19.

Zhou R, Czech B, Brennecke J, Sachidanandam R, Wohlschlegel J, Perrimon N . Processing of Drosophila endo-siRNAs depends on a specific Loquacious isoform. RNA. 2009; 15(10):1886-95. PMC: 2743050. DOI: 10.1261/rna.1611309. View

20.

Hartig J, Esslinger S, Bottcher R, Saito K, Forstemann K . Endo-siRNAs depend on a new isoform of loquacious and target artificially introduced, high-copy sequences. EMBO J. 2009; 28(19):2932-44. PMC: 2760103. DOI: 10.1038/emboj.2009.220. View