Long Double-stranded RNA Produces Specific Gene Downregulation in Giardia Lamblia

Overview

Journal J Parasitol

Specialty Parasitology

Date 2010 May 19

PMID 20476805

Citations 13

Authors

Maria R Rivero

Liudmila Kulakova

Maria C Touz

Affiliations

Soon will be listed here.

Abstract

In many eukaryotes, the introduction of double-stranded RNA (dsRNA) into cells triggers the degradation of mRNAs through a post-transcriptional gene-silencing mechanism called RNA interference or RNAi. In the present study, we found that endogenous long-dsRNA was substantially more effective at producing interference than endogenous, or exogenous, short-dsRNA expression in Giardia lamblia . The effects of this interference were not evident in the highly expressed protein tubulin or the stage-specific cyst wall protein 2. However, long-dsRNA caused potent and specific interference in the medium subunits of adaptins, the RNA-dependent RNA polymerase, and the exogenous green fluorescence protein. Our results suggest that the ability of dsRNA antisense to inhibit the expression of these specific types of proteins is indicative of a gene-specific mechanism.

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References

Touz M, Conrad J, Nash T . A novel palmitoyl acyl transferase controls surface protein palmitoylation and cytotoxicity in Giardia lamblia. Mol Microbiol. 2005; 58(4):999-1011. DOI: 10.1111/j.1365-2958.2005.04891.x. View

Marti M, Regos A, Li Y, Schraner E, Wild P, Muller N . An ancestral secretory apparatus in the protozoan parasite Giardia intestinalis. J Biol Chem. 2003; 278(27):24837-48. DOI: 10.1074/jbc.M302082200. View

Dan M, Wang A, Wang C . Inhibition of pyruvate-ferredoxin oxidoreductase gene expression in Giardia lamblia by a virus-mediated hammerhead ribozyme. Mol Microbiol. 2000; 36(2):447-56. DOI: 10.1046/j.1365-2958.2000.01863.x. View

MacRae I, Zhou K, Li F, Repic A, Brooks A, Cande W . Structural basis for double-stranded RNA processing by Dicer. Science. 2006; 311(5758):195-8. DOI: 10.1126/science.1121638. View

Rivero M, Vranych C, Bisbal M, Maletto B, Ropolo A, Touz M . Adaptor protein 2 regulates receptor-mediated endocytosis and cyst formation in Giardia lamblia. Biochem J. 2010; 428(1):33-45. PMC: 2861151. DOI: 10.1042/BJ20100096. View

Stefanic S, Morf L, Kulangara C, Regos A, Sonda S, Schraner E . Neogenesis and maturation of transient Golgi-like cisternae in a simple eukaryote. J Cell Sci. 2009; 122(Pt 16):2846-56. DOI: 10.1242/jcs.049411. View

Militello K, Refour P, Comeaux C, Duraisingh M . Antisense RNA and RNAi in protozoan parasites: working hard or hardly working?. Mol Biochem Parasitol. 2007; 157(2):117-26. DOI: 10.1016/j.molbiopara.2007.10.004. View

Ngo H, Tschudi C, Gull K, Ullu E . Double-stranded RNA induces mRNA degradation in Trypanosoma brucei. Proc Natl Acad Sci U S A. 1998; 95(25):14687-92. PMC: 24510. DOI: 10.1073/pnas.95.25.14687. View

Fire A, Xu S, Montgomery M, Kostas S, Driver S, Mello C . Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 1998; 391(6669):806-11. DOI: 10.1038/35888. View

10.

Yokota T, Sakamoto N, Enomoto N, Tanabe Y, Miyagishi M, Maekawa S . Inhibition of intracellular hepatitis C virus replication by synthetic and vector-derived small interfering RNAs. EMBO Rep. 2003; 4(6):602-8. PMC: 1319196. DOI: 10.1038/sj.embor.embor840. View

11.

Brummelkamp T, Bernards R, Agami R . A system for stable expression of short interfering RNAs in mammalian cells. Science. 2002; 296(5567):550-3. DOI: 10.1126/science.1068999. View

12.

Kaur G, Lohia A . Inhibition of gene expression with double strand RNA interference in Entamoeba histolytica. Biochem Biophys Res Commun. 2004; 320(4):1118-22. DOI: 10.1016/j.bbrc.2004.06.064. View

13.

Touz M, Kulakova L, Nash T . Adaptor protein complex 1 mediates the transport of lysosomal proteins from a Golgi-like organelle to peripheral vacuoles in the primitive eukaryote Giardia lamblia. Mol Biol Cell. 2004; 15(7):3053-60. PMC: 452563. DOI: 10.1091/mbc.e03-10-0744. View

14.

LaCount D, Bruse S, Hill K, Donelson J . Double-stranded RNA interference in Trypanosoma brucei using head-to-head promoters. Mol Biochem Parasitol. 2000; 111(1):67-76. DOI: 10.1016/s0166-6851(00)00300-5. View

15.

Prucca C, Slavin I, Quiroga R, Elias E, Rivero F, Saura A . Antigenic variation in Giardia lamblia is regulated by RNA interference. Nature. 2008; 456(7223):750-4. DOI: 10.1038/nature07585. View

16.

Hutvagner G, Mlynarova L, Nap J . Detailed characterization of the posttranscriptional gene-silencing-related small RNA in a GUS gene-silenced tobacco. RNA. 2000; 6(10):1445-54. PMC: 1370015. DOI: 10.1017/s1355838200001096. View

17.

Hannon G . RNA interference. Nature. 2002; 418(6894):244-51. DOI: 10.1038/418244a. View

18.

Touz M, Gottig N, Nash T, Lujan H . Identification and characterization of a novel secretory granule calcium-binding protein from the early branching eukaryote Giardia lamblia. J Biol Chem. 2002; 277(52):50557-63. DOI: 10.1074/jbc.M202558200. View

19.

Saraiya A, Wang C . snoRNA, a novel precursor of microRNA in Giardia lamblia. PLoS Pathog. 2008; 4(11):e1000224. PMC: 2583053. DOI: 10.1371/journal.ppat.1000224. View

20.

Nash T, Lujan H, Mowatt M, CONRAD J . Variant-specific surface protein switching in Giardia lamblia. Infect Immun. 2001; 69(3):1922-3. PMC: 98104. DOI: 10.1128/IAI.69.3.1922-1923.2001. View