title: The functional RNA domain 5BSL3.2 within the NS5B coding sequence influences hepatitis C virus IRES-mediated translation.
abstract: Hepatitis C virus (HCV) translation is mediated by an internal ribosome entry site (IRES) located at the 5' end of the genomic RNA. The 3' untranslatable region (3'UTR) stimulates translation by the recruitment of protein factors that simultaneously bind to the 5' end of the viral genome. This leads to the formation of a macromolecular complex with a closed loop conformation, similar to that described for the cap-translated mRNAs. We previously demonstrated the existence of a long-range RNA-RNA interaction involving subdomain IIId of the IRES region and the stem-loop 5BSL3.2 of the CRE element at the 3' end of the viral genome. The present study provides evidence that the enhancement of HCV IRES-dependent translation mediated by the 3'UTR is negatively controlled by the CRE region in the human hepatoma cell lines Huh-7 and Hep-G2 in a time-dependent manner. Domain 5BSL3.2 is the major partner in this process. Mutations in this motif lead to an increase in IRES activity by up to eightfold. These data support the existence of a functional high order structure in the HCV genome that involves two evolutionarily conserved RNA elements, domain IIId in the IRES and stem-loop 5BSL3.2 in the CRE region. This interaction could have a role in the circularisation of the viral genome.
authors: Instituto de Parasitología y Biomedicina López-Neyra IPBLN-CSIC, CSIC, Parque Tecnológico de Ciencias de la Salud, Avda. del Conocimiento s/n, 18100 Armilla, Granada, Spain.
authors: Cristina
authors: C
authors: Romero-López
authors: Alfredo
authors: A
authors: Berzal-Herranz
references: RNA. 2003 Mar;9(3):331-45
references: 12592007
references: J Virol. 2000 Jul;74(14):6242-50
references: 10864633
references: FEBS J. 2008 Aug;275(16):4179-97
references: 18637118
references: J Virol. 2006 Dec;80(23):11579-88
references: 16971433
references: Nucleic Acids Res. 2003 Jul 1;31(13):3406-15
references: 12824337
references: J Virol. 2001 Feb;75(3):1348-58
references: 11152508
references: Proc Natl Acad Sci U S A. 1990 Dec;87(24):9524-8
references: 2175903
references: Hepatology. 2009 Nov;50(5):1646-55
references: 19670425
references: J Gen Virol. 2006 Oct;87(Pt 10):3013-3022
references: 16963760
references: J Virol. 1993 Jun;67(6):3338-44
references: 8388503
references: J Gen Virol. 1999 Dec;80 ( Pt 12):3109-3113
references: 10567641
references: Proc Natl Acad Sci U S A. 2009 Aug 11;106(32):13517-22
references: 19628699
references: Virus Res. 2006 Jul;119(1):43-51
references: 16307817
references: Cell Mol Life Sci. 2007 Nov;64(22):2994-3006
references: 17938858
references: Virology. 2006 Feb 20;345(2):404-15
references: 16289655
references: J Virol. 2005 Jan;79(1):380-92
references: 15596831
references: J Virol. 2005 Nov;79(22):13963-73
references: 16254332
references: RNA. 2009 Aug;15(8):1528-42
references: 19541769
references: Virus Res. 2009 Nov;145(2):285-92
references: 19665505
references: J Virol. 1998 Nov;72(11):8789-96
references: 9765423
references: J Biol Chem. 2007 Jan 5;282(1):132-41
references: 17095505
references: Science. 2005 Sep 2;309(5740):1577-81
references: 16141076
references: Nucleic Acids Res. 2009 Feb;37(3):957-71
references: 19106142
references: J Virol. 2008 Sep;82(18):9008-22
references: 18614633
references: Cell. 2004 Oct 29;119(3):369-80
references: 15507208
references: Novartis Found Symp. 2006;277:23-39; discussion 40, 71-3, 251-3
references: 17319152
references: RNA. 2009 Sep;15(9):1740-52
references: 19605533
references: J Virol. 2003 Mar;77(6):3557-68
references: 12610131
references: Proc Natl Acad Sci U S A. 2004 Dec 7;101(49):16990-5
references: 15563596
references: J Virol. 2000 Nov;74(22):10430-7
references: 11044087
references: J Hum Virol. 1999 Sep-Oct;2(5):296-307
references: 10551736
references: Hepatology. 1997 Sep;26(3 Suppl 1):15S-20S
references: 9305658
references: Biol Chem. 2005 Feb;386(2):183-90
references: 15843163
references: J Virol. 2004 Oct;78(20):10865-77
references: 15452207
references: J Virol. 1992 Mar;66(3):1476-83
references: 1310759
references: Science. 1989 Apr 21;244(4902):362-4
references: 2496467
references: J Virol. 1991 Mar;65(3):1105-13
references: 1847440
references: Nat Med. 2005 Jul;11(7):791-6
references: 15951748
references: Nat Struct Biol. 2000 Dec;7(12):1105-10
references: 11101890
references: RNA. 2002 Aug;8(8):1045-55
references: 12212848
references: Nucleic Acids Res. 2009 Mar;37(4):1141-51
references: 19129232
references: Cell. 1997 Jun 13;89(6):831-8
references: 9200601
references: J Gen Virol. 2009 Jul;90(Pt 7):1659-1669
references: 19264618
references: Arch Virol. 2005 Jun;150(6):1085-99
references: 15747050
references: FEBS Lett. 2010 Feb 19;584(4):837-42
references: 20079737
references: J Biol Chem. 2005 Oct 28;280(43):36417-28
references: 16126720
references: J Virol. 2002 Jun;76(11):5326-38
references: 11991961
references: J Virol. 2000 Feb;74(4):2046-51
references: 10644379
references: RNA. 2000 Oct;6(10):1423-31
references: 11073218
references: EMBO J. 2008 Dec 17;27(24):3300-10
references: 19020517
references: J Gen Virol. 2001 Jan;82(Pt 1):113-120
references: 11125164
references: J Virol. 2003 Feb;77(3):2265-70
references: 12525663
references: Adv Virus Res. 2003;59:177-228
references: 14696330
references: Science. 1989 Apr 21;244(4902):359-62
references: 2523562
references: EMBO J. 1995 Dec 1;14(23):6010-20
references: 8846793
references: Mol Ther. 2002 Jun;5(6):676-84
references: 12027551
references: Prog Nucleic Acid Res Mol Biol. 2006;81:331-67
references: 16891176
references: Nucleic Acids Res. 2006 Mar 01;34(4):1293-303
references: 16510853
references: J Virol. 2004 Feb;78(3):1352-66
references: 14722290
references: J Virol. 2000 Dec;74(23):11347-58
references: 11070035