LSm1 binds to the Dengue virus RNA 3' UTR and is a positive regulator of Dengue virus replication

Dong,Yangchao; Yang,Jing; Ye,Wei; Wang,Yuan; Miao,Yunbo; Ding,Tianbing; Xiang,Chen; Lei,Yingfeng; Xu,Zhikai

doi:10.3892/ijmm.2015.2169

LSm1 binds to the Dengue virus RNA 3' UTR and is a positive regulator of Dengue virus replication

Authors:
- Yangchao Dong
- Jing Yang
- Wei Ye
- Yuan Wang
- Yunbo Miao
- Tianbing Ding
- Chen Xiang
- Yingfeng Lei
- Zhikai Xu
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Affiliations: Department of Microbiology, Faculty of Preclinical Medicine, The Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China, Department of Orthopaedics, Tangdu Hospital, The Fourth Military Medical University, Xi'an, Shaanxi 710038, P.R. China
Published online on: April 7, 2015 https://doi.org/10.3892/ijmm.2015.2169
Pages: 1683-1689

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Abstract

Dengue virus (DENV) is a mosquito-transmitted flavivirus that can cause severe disease in humans. The DENV positive strand RNA genome contains 5' and 3' untranslated regions (UTRs) that have been shown to be required for virus replication and interaction with host cell proteins. In the present study LSm1 was identified as a host cellular protein involved in DENV RNA replication. By using two independent methodologies, we demonstrated a critical interaction between LSm1 and the 3' UTR of DENV. Furthermore, the confocal immunofluorescence analysis showed that the interaction between LSm1 and viral RNA is located in P‑body around nucleoli in the cytoplasm. LSm1 knockdown by siRNA specifically reduced the levels of viral RNA in DENV‑infected cells and infectious DENV particles in the supernatant. These results provide evidence that LSm1 binding to the DENV RNA 3' UTR positively regulates DENV RNA replication.

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1	Wiwanitkit V: Dengue fever: diagnosis and treatment. Expert Rev Anti Infect Ther. 8:841–845. 2010. View Article : Google Scholar : PubMed/NCBI
2	Rodenhuis-Zybert IA, Wilschut J and Smit JM: Dengue virus life cycle: viral and host factors modulating infectivity. Cell Mol Life Sci. 67:2773–2786. 2010. View Article : Google Scholar : PubMed/NCBI
3	Yocupicio-Monroy M, Padmanabhan R, Medina F and del Angel RM: Mosquito La protein binds to the 3′ untranslated region of the positive and negative polarity dengue virus RNAs and relocates to the cytoplasm of infected cells. Virology. 357:29–40. 2007. View Article : Google Scholar
4	Yocupicio-Monroy RM, Medina F, Reyes-del Valle J and del Angel RM: Cellular proteins from human monocytes bind to dengue 4 virus minus-strand 3′ untranslated region RNA. J Virol. 77:3067–3076. 2003. View Article : Google Scholar : PubMed/NCBI
5	De Nova-Ocampo M, Villegas-Sepúlveda N and del Angel RM: Translation elongation factor-1alpha, La, and PTB interact with the 3′ untranslated region of dengue 4 virus RNA. Virology. 295:337–347. 2002. View Article : Google Scholar : PubMed/NCBI
6	Polacek C, Friebe P and Harris E: Poly(A)-binding protein binds to the non-polyadenylated 3′ untranslated region of dengue virus and modulates translation efficiency. J Gen Virol. 90:687–692. 2009. View Article : Google Scholar : PubMed/NCBI
7	Gomila RC, Martin GW and Gehrke L: NF90 binds the dengue virus RNA 3′ terminus and is a positive regulator of dengue virus replication. PLoS One. 6:e166872011. View Article : Google Scholar
8	Lei Y, Huang Y, Zhang H, Yu L, Zhang M and Dayton A: Functional interaction between cellular p100 and the dengue virus 3′ UTR. J Gen Virol. 92:796–806. 2011. View Article : Google Scholar
9	Paranjape SM and Harris E: Y box-binding protein-1 binds to the dengue virus 3′-untranslated region and mediates antiviral effects. J Biol Chem. 282:30497–30508. 2007. View Article : Google Scholar : PubMed/NCBI
10	Ward AM, Bidet K, Yinglin A, Ler SG, Hogue K, Blackstock W, Gunaratne J and Garcia-Blanco MA: Quantitative mass spectrometry of DENV-2 RNA-interacting proteins reveals that the DEAD-box RNA helicase DDX6 binds the DB1 and DB2 3′ UTR structures. RNA Biol. 8:1173–1186. 2011. View Article : Google Scholar : PubMed/NCBI
11	Beggs JD: Lsm proteins and RNA processing. Biochem Soc Trans. 33:433–438. 2005. View Article : Google Scholar : PubMed/NCBI
12	Khusial P, Plaag R and Zieve GW: LSm proteins form heptameric rings that bind to RNA via repeating motifs. Trends Biochem Sci. 30:522–528. 2005. View Article : Google Scholar : PubMed/NCBI
13	Diez J, Ishikawa M, Kaido M and Ahlquist P: Identification and characterization of a host protein required for efficient template selection in viral RNA replication. Proc Natl Acad Sci USA. 97:3913–3918. 2000. View Article : Google Scholar : PubMed/NCBI
14	Galao RP, Chari A, Alves-Rodrigues I, Lobão D, Mas A, Kambach C, Fischer U and Díez J: LSm1-7 complexes bind to specific sites in viral RNA genomes and regulate their translation and replication. RNA. 16:817–827. 2010. View Article : Google Scholar : PubMed/NCBI
15	Pérez-Vilaró G, Scheller N, Saludes V and Díez J: Hepatitis C virus infection alters P-body composition but is independent of P-body granules. J Virol. 86:8740–8749. 2012. View Article : Google Scholar : PubMed/NCBI
16	Scheller N, Mina LB, Galao RP, Chari A, Giménez-Barcons M, Noueiry A, Fischer U, Meyerhans A and Díez J: Translation and replication of hepatitis C virus genomic RNA depends on ancient cellular proteins that control mRNA fates. Proc Natl Acad Sci USA. 106:13517–13522. 2009. View Article : Google Scholar : PubMed/NCBI
17	Pang X, Zhang M and Dayton AI: Development of Dengue virus type 2 replicons capable of prolonged expression in host cells. BMC Microbiol. 1:182001. View Article : Google Scholar : PubMed/NCBI
18	Polo S, Ketner G, Levis R and Falgout B: Infectious RNA transcripts from full-length dengue virus type 2 cDNA clones made in yeast. J Virol. 71:5366–5374. 1997.PubMed/NCBI
19	Emara MM and Brinton MA: Interaction of TIA-1/TIAR with West Nile and dengue virus products in infected cells interferes with stress granule formation and processing body assembly. Proc Natl Acad Sci USA. 104:9041–9046. 2007. View Article : Google Scholar : PubMed/NCBI
20	Eulalio A, Behm-Ansmant I, Schweizer D and Izaurralde E: P-body formation is a consequence, not the cause, of RNA- mediated gene silencing. Mol Cell Biol. 27:3970–3981. 2007. View Article : Google Scholar : PubMed/NCBI
21	Jakymiw A, Pauley KM, Li S, Ikeda K, Lian S, Eystathioy T, Satoh M, Fritzler MJ and Chan EK: The role of GW/P-bodies in RNA processing and silencing. J Cell Sci. 120:1317–1323. 2007. View Article : Google Scholar : PubMed/NCBI
22	Chan EK, Yao B and Fritzler MJ: Reflections on ten years of history of, and future prospects for, GW182 and GW/P body research. Adv Exp Med Biol. 768:261–270. 2013. View Article : Google Scholar
23	Coller J and Parker R: Eukaryotic mRNA decapping. Annu Rev Biochem. 73:861–890. 2004. View Article : Google Scholar : PubMed/NCBI
24	Yu SF, Lujan P, Jackson DL, Emerman M and Linial ML: The DEAD-box RNA helicase DDX6 is required for efficient encapsidation of a retroviral genome. PLoS Pathog. 7:e10023032011. View Article : Google Scholar : PubMed/NCBI
25	Ariumi Y, Kuroki M, Kushima Y, Osugi K, Hijikata M, Maki M, Ikeda M and Kato N: Hepatitis C virus hijacks P-body and stress granule components around lipid droplets. J Virol. 85:6882–6892. 2011. View Article : Google Scholar : PubMed/NCBI
26	Men R, Bray M, Clark D, Chanock RM and Lai CJ: Dengue type 4 virus mutants containing deletions in the 3′ noncoding region of the RNA genome: analysis of growth restriction in cell culture and altered viremia pattern and immunogenicity in rhesus monkeys. J Virol. 70:3930–3937. 1996.PubMed/NCBI
27	Agis-Juárez RA, Galván I, Medina F, Daikoku T, Padmanabhan R, Ludert JE and del Angel RM: Polypyrimidine tract-binding protein is relocated to the cytoplasm and is required during dengue virus infection in Vero cells. J Gen Virol. 90:2893–2901. 2009. View Article : Google Scholar : PubMed/NCBI
28	Roberts AP, Doidge R, Tarr AW and Jopling CL: 2013. The P body protein LSm1 contributes to stimulation of hepatitis C virus translation, but not replication, by microRNA-122. Nucleic Acids Res. 42:1257–1269. 2014. View Article : Google Scholar :
29	Chowdhury A, Mukhopadhyay J and Tharun S: The decapping activator Lsm1p-7p-Pat1p complex has the intrinsic ability to distinguish between oligoadenylated and polyadenylated RNAs. RNA. 13:998–1016. 2007. View Article : Google Scholar : PubMed/NCBI
30	Beckham CJ and Parker R: P bodies, stress granules, and viral life cycles. Cell Host Microbe. 3:206–212. 2008. View Article : Google Scholar : PubMed/NCBI
31	Parker R and Sheth U: P bodies and the control of mRNA translation and degradation. Mol Cell. 25:635–646. 2007. View Article : Google Scholar : PubMed/NCBI
32	Huys A, Thibault PA and Wilson JA: Modulation of hepatitis C virus RNA accumulation and translation by DDX6 and miR-122 are mediated by separate mechanisms. PLoS One. 8:e674372013. View Article : Google Scholar : PubMed/NCBI