title: Binding of cellular proteins to the leader RNA of equine arteritis virus.
abstract: The genome of equine arteritis virus (EAV) produces a 3' coterminal-nested set of six subgenomic (sg) viral RNAs during virus replication cycle, and each set possesses a common leader sequence of 206 nucleotides (nt) in length derived from the 5' end of the viral genome. Given the presence of the leader region within both genomic and sg mRNAs, it is likely to contain cis-acting signals that may interact with cellular or viral proteins for RNA synthesis. Gel mobility shift assays indicated that proteins in Vero cell cytoplasmic extracts formed complexes with the positive (+) and negative (-) strands of the EAV leader RNA. Several cell proteins with molecular masses ranging from 74 to 31 kDa and 58 to 32 kDa were detected in UV-induced cross-linking assays with the EAV leader RNA (+) and (-) strands, respectively. In both cases, intense bands were observed at the 58-52 kDa molecular weight markers. Results from competition gel mobility shift assays using overlapping cold RNA probes spanning the leader RNA (+) strand indicated that nt 140-206 are not necessary for binding to cell proteins.
authors.affiliation: Department of Biological Sciences, University of Québec at Montréal, P.O. Box 8888, Succursale Centre-Ville, Montréal, Québec, H3C 3P8, Canada. archambault.denis@uqam.ca
authors.forename: Denis
authors.initials: D
authors.lastname: Archambault
authors.forename: Marie-Claude
authors.initials: MC
authors.lastname: St-Louis
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Martin
mesh_terms: D020121:5' Untranslated Regions / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D007240:Equartevirus / Q000235:genetics / Q000378:metabolism*; D008970:Molecular Weight; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D015347:RNA Probes
chemical_list: D020121:5' Untranslated Regions; D011506:Proteins; D015347:RNA Probes
references.citation: J Gen Virol. 1998 May;79 ( Pt 5):961-79
references.pmid: 9603311
references.citation: Proc Natl Acad Sci U S A. 1997 Mar 18;94(6):2249-54
references.pmid: 9122180
references.citation: J Virol. 1995 Apr;69(4):2016-23
references.pmid: 7884846
references.citation: J Virol. 2000 Mar;74(5):2219-26
references.pmid: 10666252
references.citation: J Virol. 1999 Jan;73(1):772-7
references.pmid: 9847386
references.citation: Biotechniques. 1999 Nov;27(5):1032-9, 1042
references.pmid: 10572651
references.citation: J Virol. 1993 Aug;67(8):5003-11
references.pmid: 8392625
references.citation: J Gen Virol. 2002 Apr;83(Pt 4):795-800
references.pmid: 11907328
references.citation: Virology. 1998 Apr 25;244(1):1-12
references.pmid: 9581772
references.citation: RNA. 2004 Mar;10(3):424-37
references.pmid: 14970388
references.citation: J Gen Virol. 2003 Sep;84(Pt 9):2317-22
references.pmid: 12917451
references.citation: Proc Natl Acad Sci U S A. 1999 Oct 12;96(21):12056-61
references.pmid: 10518575
references.citation: J Virol. 1998 May;72(5):4341-51
references.pmid: 9557724
references.citation: Proc Natl Acad Sci U S A. 1997 Sep 2;94(18):9544-9
references.pmid: 9275159
references.citation: Adv Exp Med Biol. 1995;380:499-506
references.pmid: 8830530
references.citation: Clin Diagn Lab Immunol. 2002 May;9(3):698-703
references.pmid: 11986280
references.citation: J Virol. 1993 Dec;67(12):7215-22
references.pmid: 8230443
references.citation: Can J Vet Res. 1998 Jul;62(3):224-30
references.pmid: 9684053