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Translation elongation factor-1alpha, La, and PTB interact with the 3' untranslated region of dengue 4 virus RNA.
Virology. 2002 Apr 10; 295(2):337-47.V

Abstract

The 384-nt long 3' untranslated region (3'UTR) of dengue 4 virus (DEN4) is not polyadenylated, but contains the adjacent thermodynamically stable conserved short and long stem-loop structures (L-SL) and the conserved sequences CS1 and CS2. The latter are duplicated (CS2A and CS2B) in DEN4. Dengue virus replication, like that of other RNA viruses, might involve the cis-elements located within the 3'UTR and the trans-acting factors that could interact with the viral replicase to function as a replicase complex. The identification and characterization of viral and cellular proteins involved in the interaction with the 3'UTR of dengue virus will help us to understand the cellular requirements for viral replication. To determine these requirements, mobility shift and cross-linking assays were performed with uninfected and DEN4-infected C6/36 cell extracts as well as the different segments of the 3'UTR. Our results revealed that RNA-protein complexes were formed with the RNAs which involved the domains CS2A, CS2B, CS1, and L-SL. The minimum RNA sequence that was able to form specific and stable complexes with cellular proteins was the CS1-L-SL region. Using UV-induced cross-linking we identified eight proteins with molecular weights of 34, 39, 51, 52, 56, 62, 72, and 84 kDa that bound to the complete 3'UTR. The translation elongation factor-1alpha (EF-1alpha) bound to the complete 3'UTR and to the CS1-L-SL region. In addition, the recombinant GST-human La autoantigen bound to the 3'UTR and to the CS1-L-SL region as demonstrated by mobility shift and cross-linking assays. Although different antibodies against PTB were unable to react with any of the cellular proteins from C6/36, the recombinant His-PTB protein did bind to the complete 3'UTR and to the CS1-L-SL region. The specific binding of La and PTB to the sequences considered essential for viral RNA replication may suggest that these proteins could function as RNA chaperones to maintain RNA structure in a conformation that favors viral replication, while EF-1alpha may function as an RNA helicase.

Authors+Show Affiliations

Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del IPN, Mexico City 07360, Mexico.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12033793

Citation

De Nova-Ocampo, Mónica, et al. "Translation Elongation Factor-1alpha, La, and PTB Interact With the 3' Untranslated Region of Dengue 4 Virus RNA." Virology, vol. 295, no. 2, 2002, pp. 337-47.
De Nova-Ocampo M, Villegas-Sepúlveda N, del Angel RM. Translation elongation factor-1alpha, La, and PTB interact with the 3' untranslated region of dengue 4 virus RNA. Virology. 2002;295(2):337-47.
De Nova-Ocampo, M., Villegas-Sepúlveda, N., & del Angel, R. M. (2002). Translation elongation factor-1alpha, La, and PTB interact with the 3' untranslated region of dengue 4 virus RNA. Virology, 295(2), 337-47.
De Nova-Ocampo M, Villegas-Sepúlveda N, del Angel RM. Translation Elongation Factor-1alpha, La, and PTB Interact With the 3' Untranslated Region of Dengue 4 Virus RNA. Virology. 2002 Apr 10;295(2):337-47. PubMed PMID: 12033793.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Translation elongation factor-1alpha, La, and PTB interact with the 3' untranslated region of dengue 4 virus RNA. AU - De Nova-Ocampo,Mónica, AU - Villegas-Sepúlveda,Nicolás, AU - del Angel,Rosa M, PY - 2002/5/30/pubmed PY - 2002/7/13/medline PY - 2002/5/30/entrez SP - 337 EP - 47 JF - Virology JO - Virology VL - 295 IS - 2 N2 - The 384-nt long 3' untranslated region (3'UTR) of dengue 4 virus (DEN4) is not polyadenylated, but contains the adjacent thermodynamically stable conserved short and long stem-loop structures (L-SL) and the conserved sequences CS1 and CS2. The latter are duplicated (CS2A and CS2B) in DEN4. Dengue virus replication, like that of other RNA viruses, might involve the cis-elements located within the 3'UTR and the trans-acting factors that could interact with the viral replicase to function as a replicase complex. The identification and characterization of viral and cellular proteins involved in the interaction with the 3'UTR of dengue virus will help us to understand the cellular requirements for viral replication. To determine these requirements, mobility shift and cross-linking assays were performed with uninfected and DEN4-infected C6/36 cell extracts as well as the different segments of the 3'UTR. Our results revealed that RNA-protein complexes were formed with the RNAs which involved the domains CS2A, CS2B, CS1, and L-SL. The minimum RNA sequence that was able to form specific and stable complexes with cellular proteins was the CS1-L-SL region. Using UV-induced cross-linking we identified eight proteins with molecular weights of 34, 39, 51, 52, 56, 62, 72, and 84 kDa that bound to the complete 3'UTR. The translation elongation factor-1alpha (EF-1alpha) bound to the complete 3'UTR and to the CS1-L-SL region. In addition, the recombinant GST-human La autoantigen bound to the 3'UTR and to the CS1-L-SL region as demonstrated by mobility shift and cross-linking assays. Although different antibodies against PTB were unable to react with any of the cellular proteins from C6/36, the recombinant His-PTB protein did bind to the complete 3'UTR and to the CS1-L-SL region. The specific binding of La and PTB to the sequences considered essential for viral RNA replication may suggest that these proteins could function as RNA chaperones to maintain RNA structure in a conformation that favors viral replication, while EF-1alpha may function as an RNA helicase. SN - 0042-6822 UR - https://www.unboundmedicine.com/medline/citation/12033793/Translation_elongation_factor_1alpha_La_and_PTB_interact_with_the_3'_untranslated_region_of_dengue_4_virus_RNA_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0042682202914078 DB - PRIME DP - Unbound Medicine ER -