title: Expression and processing of a small nucleolar RNA from the Epstein-Barr virus genome.
abstract: Small nucleolar RNAs (snoRNAs) are localized within the nucleolus, a sub-nuclear compartment, in which they guide ribosomal or spliceosomal RNA modifications, respectively. Up until now, snoRNAs have only been identified in eukaryal and archaeal genomes, but are notably absent in bacteria. By screening B lymphocytes for expression of non-coding RNAs (ncRNAs) induced by the Epstein-Barr virus (EBV), we here report, for the first time, the identification of a snoRNA gene within a viral genome, designated as v-snoRNA1. This genetic element displays all hallmark sequence motifs of a canonical C/D box snoRNA, namely C/C'- as well as D/D'-boxes. The nucleolar localization of v-snoRNA1 was verified by in situ hybridisation of EBV-infected cells. We also confirmed binding of the three canonical snoRNA proteins, fibrillarin, Nop56 and Nop58, to v-snoRNA1. The C-box motif of v-snoRNA1 was shown to be crucial for the stability of the viral snoRNA; its selective deletion in the viral genome led to a complete down-regulation of v-snoRNA1 expression levels within EBV-infected B cells. We further provide evidence that v-snoRNA1 might serve as a miRNA-like precursor, which is processed into 24 nt sized RNA species, designated as v-snoRNA1(24pp). A potential target site of v-snoRNA1(24pp) was identified within the 3'-UTR of BALF5 mRNA which encodes the viral DNA polymerase. V-snoRNA1 was found to be expressed in all investigated EBV-positive cell lines, including lymphoblastoid cell lines (LCL). Interestingly, induction of the lytic cycle markedly up-regulated expression levels of v-snoRNA1 up to 30-fold. By a computational approach, we identified a v-snoRNA1 homolog in the rhesus lymphocryptovirus genome. This evolutionary conservation suggests an important role of v-snoRNA1 during gamma-herpesvirus infection.
authors.affiliation: Innsbruck Biocenter, Division of Genomics and RNomics, Innsbruck Medical University, Innsbruck, Austria.
authors.forename: Roland
authors.initials: R
authors.lastname: Hutzinger
authors.forename: Regina
authors.initials: R
authors.lastname: Feederle
authors.forename: Jan
authors.initials: J
authors.lastname: Mrazek
authors.forename: Natalia
authors.initials: N
authors.lastname: Schiefermeier
authors.forename: Piotr J
authors.initials: PJ
authors.lastname: Balwierz
authors.forename: Mihaela
authors.initials: M
authors.lastname: Zavolan
authors.forename: Norbert
authors.initials: N
authors.lastname: Polacek
authors.forename: Henri-Jacques
authors.initials: HJ
authors.lastname: Delecluse
authors.forename: Alexander
authors.initials: A
authors.lastname: Hüttenhofer
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000821:virology; D001483:Base Sequence; D045744:Cell Line, Tumor; D015967:Gene Expression Regulation, Viral; D016679:Genome, Viral; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008253:Macaca mulatta; D008969:Molecular Sequence Data; D020537:RNA, Small Nucleolar / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics
chemical_list: D020537:RNA, Small Nucleolar; D012367:RNA, Viral
references.citation: EMBO J. 2001 Jul 16;20(14):3617-22
references.pmid: 11447102
references.citation: Proc Natl Acad Sci U S A. 2007 Oct 9;104(41):16164-9
references.pmid: 17911266
references.citation: EMBO J. 2001 Jun 1;20(11):2943-53
references.pmid: 11387227
references.citation: Genome Res. 2002 Nov;12(11):1739-48
references.pmid: 12421761
references.citation: EMBO J. 1990 Dec;9(13):4503-9
references.pmid: 2265615
references.citation: Science. 2006 Jan 13;311(5758):230-2
references.pmid: 16357227
references.citation: J Virol. 2009 Apr;83(7):3333-41
references.pmid: 19144710
references.citation: Proc Natl Acad Sci U S A. 1972 Feb;69(2):383-7
references.pmid: 4333982
references.citation: J Virol. 1990 Jun;64(6):2759-67
references.pmid: 2159542
references.citation: Nat Rev Mol Cell Biol. 2007 Mar;8(3):209-20
references.pmid: 17318225
references.citation: Nature. 2001 Jan 18;409(6818):363-6
references.pmid: 11201747
references.citation: Biochimie. 1995;77(1-2):22-9
references.pmid: 7599273
references.citation: Nucleic Acids Res. 2004 Aug 27;32(15):4646-56
references.pmid: 15333696
references.citation: EMBO J. 2000 Jun 15;19(12):3080-9
references.pmid: 10856251
references.citation: RNA. 2007 Jun;13(6):930-6
references.pmid: 17456563
references.citation: Science. 2004 Apr 30;304(5671):734-6
references.pmid: 15118162
references.citation: Mol Cell Biol. 2004 May;24(10):4522-33
references.pmid: 15121869
references.citation: Nat Rev Genet. 2008 Jul;9(7):503-7
references.pmid: 18490927
references.citation: J Virol. 1998 Apr;72(4):2969-74
references.pmid: 9525618
references.citation: Science. 2008 Mar 28;319(5871):1787-9
references.pmid: 18369136
references.citation: Nucleic Acids Res. 2007;35(3):962-74
references.pmid: 17259222
references.citation: Cell. 1988 Nov 4;55(3):427-33
references.pmid: 2846181
references.citation: Science. 2000 Apr 21;288(5465):517-22
references.pmid: 10775111
references.citation: Proc Natl Acad Sci U S A. 1998 Jul 7;95(14):8245-50
references.pmid: 9653172
references.citation: Trends Genet. 2005 May;21(5):289-97
references.pmid: 15851066
references.citation: J Virol. 1981 Oct;40(1):11-9
references.pmid: 6270367
references.citation: J Virol. 2007 Sep;81(18):9967-75
references.pmid: 17626073
references.citation: Mol Cell Biol. 1992 Oct;12(10):4456-63
references.pmid: 1406634
references.citation: Science. 2004 Apr 23;304(5670):594-6
references.pmid: 15105502
references.citation: PLoS Pathog. 2006 Mar;2(3):e23
references.pmid: 16557291
references.citation: Cell. 1996 Jun 28;85(7):1077-88
references.pmid: 8674114
references.citation: Nucleic Acids Res. 2008 May;36(8):2677-89
references.pmid: 18346967
references.citation: Curr Opin Microbiol. 2006 Apr;9(2):229-36
references.pmid: 16529986
references.citation: Methods Mol Biol. 2005;292:353-70
references.pmid: 15507720
references.citation: Virology. 2009 Apr 10;386(2):387-97
references.pmid: 19217135
references.citation: Cell. 2002 Apr 19;109(2):145-8
references.pmid: 12007400
references.citation: Exp Cell Res. 1965 Mar;37:614-36
references.pmid: 14315085
references.citation: Proc Natl Acad Sci U S A. 2000 Aug 1;97(16):8955-60
references.pmid: 10922055
references.citation: J Virol. 2007 Jan;81(2):1033-6
references.pmid: 17079300
references.citation: J Virol. 2007 Jun;81(12):6389-401
references.pmid: 17409162
references.citation: J Virol. 2009 May;83(10):4952-62
references.pmid: 19264771
references.citation: Nucleic Acids Res. 2008 Feb;36(2):666-75
references.pmid: 18073197
references.citation: Nature. 2002 Jun 27;417(6892):967-70
references.pmid: 12068309
references.citation: Mol Cell. 2008 Nov 21;32(4):519-28
references.pmid: 19026782
references.citation: J Cell Biol. 2006 May 8;173(3):319-25
references.pmid: 16682524
references.citation: Gene. 1995 May 26;158(1):9-14
references.pmid: 7789817
references.citation: Cell. 2005 Nov 18;123(4):631-40
references.pmid: 16271387
references.citation: J Exp Med. 2008 Oct 27;205(11):2551-60
references.pmid: 18838543
references.citation: Curr Biol. 2005 May 24;15(10):974-9
references.pmid: 15916956
references.citation: EMBO J. 1998 Jul 1;17(13):3747-57
references.pmid: 9649444
references.citation: Nucleic Acids Res. 1998 Apr 1;26(7):1576-87
references.pmid: 9512526
references.citation: Cell. 2007 Jun 29;129(7):1401-14
references.pmid: 17604727
references.citation: PLoS Pathog. 2008 Nov;4(11):e1000224
references.pmid: 19043559
references.citation: Cell. 1997 May 30;89(5):799-809
references.pmid: 9182768
references.citation: Annu Rev Immunol. 2007;25:587-617
references.pmid: 17378764
references.citation: Oncologist. 2003;8(1):83-98
references.pmid: 12604735
references.citation: Nucleic Acids Res. 2007;35(10):e73
references.pmid: 17478510
references.citation: Oncogene. 2006 Oct 9;25(46):6211-9
references.pmid: 17028601
references.citation: J Virol. 2005 Feb;79(3):1724-33
references.pmid: 15650197
references.citation: Int J Cancer. 1986 Sep 15;38(3):425-32
references.pmid: 3017870
references.citation: Science. 2002 Sep 20;297(5589):2053-6
references.pmid: 12242443
references.citation: Nat Rev Genet. 2004 Apr;5(4):316-23
references.pmid: 15131654
references.citation: Proc Natl Acad Sci U S A. 1981 Feb;78(2):805-9
references.pmid: 6262773
references.citation: Curr Opin Chem Biol. 2002 Dec;6(6):835-43
references.pmid: 12470739
references.citation: Nat Rev Genet. 2006 Jun;7(6):475-82
references.pmid: 16622413
references.citation: J Virol. 2008 Sep;82(18):9094-106
references.pmid: 18614630
references.citation: Nat Med. 2002 Oct;8(10):1098-104
references.pmid: 12219084
references.citation: J Virol. 2000 Nov;74(21):10142-52
references.pmid: 11024143
references.citation: Mol Cell Biol. 2004 Jul;24(13):5797-807
references.pmid: 15199136
references.citation: Biochimie. 2002 Aug;84(8):775-90
references.pmid: 12457565
references.citation: Expert Rev Mol Med. 2001 Nov 15;3(28):1-20
references.pmid: 14585152
references.citation: Proc Natl Acad Sci U S A. 1973 Jan;70(1):190-4
references.pmid: 4346033