PMID- 24825891 OWN - NLM STAT- MEDLINE DCOM- 20140902 LR - 20211021 IS - 1091-6490 (Electronic) IS - 0027-8424 (Print) IS - 0027-8424 (Linking) VI - 111 IP - 21 DP - 2014 May 27 TI - Transactivation of programmed ribosomal frameshifting by a viral protein. PG - E2172-81 LID - 10.1073/pnas.1321930111 [doi] AB - Programmed -1 ribosomal frameshifting (-1 PRF) is a widely used translational mechanism facilitating the expression of two polypeptides from a single mRNA. Commonly, the ribosome interacts with an mRNA secondary structure that promotes -1 frameshifting on a homopolymeric slippery sequence. Recently, we described an unusual -2 frameshifting (-2 PRF) signal directing efficient expression of a transframe protein [nonstructural protein 2TF (nsp2TF)] of porcine reproductive and respiratory syndrome virus (PRRSV) from an alternative reading frame overlapping the viral replicase gene. Unusually, this arterivirus PRF signal lacks an obvious stimulatory RNA secondary structure, but as confirmed here, can also direct the occurrence of -1 PRF, yielding a third, truncated nsp2 variant named "nsp2N." Remarkably, we now show that both -2 and -1 PRF are transactivated by a protein factor, specifically a PRRSV replicase subunit (nsp1beta). Embedded in nsp1beta's papain-like autoproteinase domain, we identified a highly conserved, putative RNA-binding motif that is critical for PRF transactivation. The minimal RNA sequence required for PRF was mapped within a 34-nt region that includes the slippery sequence and a downstream conserved CCCANCUCC motif. Interaction of nsp1beta with the PRF signal was demonstrated in pull-down assays. These studies demonstrate for the first time, to our knowledge, that a protein can function as a transactivator of ribosomal frameshifting. The newly identified frameshifting determinants provide potential antiviral targets for arterivirus disease control and prevention. Moreover, protein-induced transactivation of frameshifting may be a widely used mechanism, potentially including previously undiscovered viral strategies to regulate viral gene expression and/or modulate host cell translation upon infection. FAU - Li, Yanhua AU - Li Y AD - Department of Veterinary and Biomedical Sciences and Department of Biology/Microbiology, South Dakota State University, Brookings, SD 57007;Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506; FAU - Treffers, Emmely E AU - Treffers EE AD - Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands;Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands; FAU - Napthine, Sawsan AU - Napthine S AD - Division of Virology, Department of Pathology, University of Cambridge, Cambridge CB2 1QP, United Kingdom; and. FAU - Tas, Ali AU - Tas A AD - Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands; FAU - Zhu, Longchao AU - Zhu L AD - Department of Veterinary and Biomedical Sciences and Department of Biology/Microbiology, South Dakota State University, Brookings, SD 57007;Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506; FAU - Sun, Zhi AU - Sun Z AD - Department of Veterinary and Biomedical Sciences and Department of Biology/Microbiology, South Dakota State University, Brookings, SD 57007; FAU - Bell, Susanne AU - Bell S AD - Division of Virology, Department of Pathology, University of Cambridge, Cambridge CB2 1QP, United Kingdom; and. FAU - Mark, Brian L AU - Mark BL AD - Department of Microbiology, University of Manitoba, Winnipeg, MB, Canada R3T 2N2. FAU - van Veelen, Peter A AU - van Veelen PA AD - Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands; FAU - van Hemert, Martijn J AU - van Hemert MJ AD - Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands; FAU - Firth, Andrew E AU - Firth AE AD - Division of Virology, Department of Pathology, University of Cambridge, Cambridge CB2 1QP, United Kingdom; and. FAU - Brierley, Ian AU - Brierley I AD - Division of Virology, Department of Pathology, University of Cambridge, Cambridge CB2 1QP, United Kingdom; and yfang@vet.k-state.edu ib103@mole.bio.cam.ac.uk e.j.snijder@lumc.nl. FAU - Snijder, Eric J AU - Snijder EJ AD - Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands; yfang@vet.k-state.edu ib103@mole.bio.cam.ac.uk e.j.snijder@lumc.nl. FAU - Fang, Ying AU - Fang Y AD - Department of Veterinary and Biomedical Sciences and Department of Biology/Microbiology, South Dakota State University, Brookings, SD 57007;Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506; yfang@vet.k-state.edu ib103@mole.bio.cam.ac.uk e.j.snijder@lumc.nl. LA - eng GR - Wellcome Trust/United Kingdom GR - 088789/WT_/Wellcome Trust/United Kingdom GR - BB/G008205/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20140513 PL - United States TA - Proc Natl Acad Sci U S A JT - Proceedings of the National Academy of Sciences of the United States of America JID - 7505876 RN - 0 (Rosaniline Dyes) RN - 0 (Viral Nonstructural Proteins) RN - EC 1.13.12.- (Luciferases) RN - M1ZRX790SI (coomassie Brilliant Blue) SB - IM MH - Animals MH - Cell Line MH - Chromatography, Liquid MH - Electrophoresis, Polyacrylamide Gel MH - Frameshifting, Ribosomal/*physiology MH - Gene Expression Regulation, Viral/*genetics MH - HEK293 Cells MH - Haplorhini MH - Humans MH - Immunoassay MH - Luciferases MH - Porcine respiratory and reproductive syndrome virus/*genetics MH - Rosaniline Dyes MH - Tandem Mass Spectrometry MH - Transcriptional Activation/*physiology MH - Viral Nonstructural Proteins/*physiology PMC - PMC4040542 OTO - NOTNLM OT - genetic recoding OT - nsp1beta OT - translational control COIS- Conflict of interest statement: The authors have filed a patent application that relates to some aspects of this work. EDAT- 2014/05/16 06:00 MHDA- 2014/09/03 06:00 PMCR- 2014/11/27 CRDT- 2014/05/15 06:00 PHST- 2014/05/15 06:00 [entrez] PHST- 2014/05/16 06:00 [pubmed] PHST- 2014/09/03 06:00 [medline] PHST- 2014/11/27 00:00 [pmc-release] AID - 1321930111 [pii] AID - 201321930 [pii] AID - 10.1073/pnas.1321930111 [doi] PST - ppublish SO - Proc Natl Acad Sci U S A. 2014 May 27;111(21):E2172-81. doi: 10.1073/pnas.1321930111. Epub 2014 May 13.