{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T09:38:19Z","timestamp":1773308299372,"version":"3.50.1"},"reference-count":49,"publisher":"Microbiology Society","issue":"6","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2007,6,1]]},"abstract":"<jats:p>Vaccinia virus (VACV) encodes many immunomodulatory proteins, including inhibitors of apoptosis and modulators of innate immune signalling. VACV protein N1 is an intracellular homodimer that contributes to virus virulence and was reported to inhibit nuclear factor (NF)-<jats:italic>\u03ba<\/jats:italic>B signalling. However, analysis of NF-<jats:italic>\u03ba<\/jats:italic>B signalling in cells infected with recombinant viruses with or without the <jats:italic>N1L<\/jats:italic> gene showed no difference in NF-<jats:italic>\u03ba<\/jats:italic>B-dependent gene expression. Given that N1 promotes virus virulence, other possible functions of N1 were investigated and this revealed that N1 is an inhibitor of apoptosis in cells transfected with the <jats:italic>N1L<\/jats:italic> gene and in the context of VACV infection. In support of this finding virally expressed N1 co-precipitated with endogenous pro-apoptotic Bcl-2 proteins Bid, Bad and Bax as well as with Bad and Bax expressed by transfection. In addition, the crystal structure of N1 was solved to 2.9\u2005\u00c5 resolution (0.29\u2005nm). Remarkably, although N1 shows no sequence similarity to cellular proteins, its three-dimensional structure closely resembles Bcl-x<jats:sub>L<\/jats:sub> and other members of the Bcl-2 protein family. The structure also reveals that N1 has a constitutively open surface groove similar to the grooves of other anti-apoptotic Bcl-2 proteins, which bind the BH3 motifs of pro-apoptotic Bcl-2 family members. Molecular modelling of BH3 peptides into the N1 surface groove, together with analysis of their physico-chemical properties, suggests a mechanism for the specificity of peptide recognition. This study illustrates the importance of the evolutionary conservation of structure, rather than sequence, in protein function and reveals a novel anti-apoptotic protein from orthopoxviruses.<\/jats:p>","DOI":"10.1099\/vir.0.82772-0","type":"journal-article","created":{"date-parts":[[2007,5,7]],"date-time":"2007-05-07T17:28:46Z","timestamp":1178558926000},"page":"1656-1666","source":"Crossref","is-referenced-by-count":152,"title":["Functional and structural studies of the vaccinia virus virulence factor N1 reveal a Bcl-2-like anti-apoptotic protein"],"prefix":"10.1099","volume":"88","author":[{"given":"Samantha","family":"Cooray","sequence":"first","affiliation":[{"name":"Department of Virology, Faculty of Medicine, Imperial College London, St Mary's Campus, Norfolk Place, London W2 1PG, UK"}]},{"given":"Mohammad W.","family":"Bahar","sequence":"additional","affiliation":[{"name":"The Oxford Protein Production Facility and The Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK"}]},{"given":"Nicola G. A.","family":"Abrescia","sequence":"additional","affiliation":[{"name":"The Oxford Protein Production Facility and The Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK"}]},{"given":"Colin E.","family":"McVey","sequence":"additional","affiliation":[{"name":"Department of Virology, Faculty of Medicine, Imperial College London, St Mary's Campus, Norfolk Place, London W2 1PG, UK"}]},{"given":"Nathan W.","family":"Bartlett","sequence":"additional","affiliation":[{"name":"Department of Virology, Faculty of Medicine, Imperial College London, St Mary's Campus, Norfolk Place, London W2 1PG, UK"}]},{"given":"Ron A.-J.","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Virology, Faculty of Medicine, Imperial College London, St Mary's Campus, Norfolk Place, London W2 1PG, UK"}]},{"given":"David I.","family":"Stuart","sequence":"additional","affiliation":[{"name":"The Oxford Protein Production Facility and The Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK"}]},{"given":"Jonathan M.","family":"Grimes","sequence":"additional","affiliation":[{"name":"The Oxford Protein Production Facility and The Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK"}]},{"given":"Geoffrey L.","family":"Smith","sequence":"additional","affiliation":[{"name":"Department of Virology, Faculty of Medicine, Imperial College London, St Mary's Campus, Norfolk Place, London W2 1PG, UK"}]}],"member":"345","reference":[{"key":"R1","doi-asserted-by":"crossref","first-page":"3815","DOI":"10.1128\/JVI.74.8.3815-3831.2000","article-title":"The genome of fowlpox virus","volume":"74","author":"Afonso","year":"2000","journal-title":"J Virol"},{"key":"R2","doi-asserted-by":"crossref","first-page":"971","DOI":"10.1128\/JVI.75.2.971-978.2001","article-title":"The genome of turkey herpesvirus","volume":"75","author":"Afonso","year":"2001","journal-title":"J Virol"},{"key":"R3","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1110\/ps.062454707","article-title":"Vaccinia virus N1L protein resembles a B cell lymphoma-2 (Bcl-2) family protein","volume":"16","author":"Aoyagi","year":"2007","journal-title":"Protein Sci"},{"key":"R4","doi-asserted-by":"crossref","first-page":"1965","DOI":"10.1099\/0022-1317-83-8-1965","article-title":"The vaccinia virus N1L protein is an intracellular homodimer that promotes virulence","volume":"83","author":"Bartlett","year":"2002","journal-title":"J Gen Virol"},{"key":"R5","doi-asserted-by":"crossref","first-page":"1207","DOI":"10.1038\/sj.emboj.7600104","article-title":"Phosphorylation of BCL-2 regulates ER Ca2+ homeostasis and apoptosis","volume":"23","author":"Bassik","year":"2004","journal-title":"EMBO J"},{"key":"R6","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1107\/S0021889803002012","article-title":"A procedure for setting up high-throughput nanolitre crystallization experiments. II. Crystallization results","volume":"36","author":"Brown","year":"2003","journal-title":"J Appl Crystallogr"},{"key":"R7","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1107\/S0907444998003254","article-title":"Crystallography & NMR system: a new software suite for macromolecular structure determination","volume":"54","author":"Br\u00fcnger","year":"1998","journal-title":"Acta Crystallogr D Biol Crystallogr"},{"key":"R8","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1016\/S0092-8674(00)80572-3","article-title":"Solution structure of BID, an intracellular amplifier of apoptotic signaling","volume":"96","author":"Chou","year":"1999","journal-title":"Cell"},{"key":"R9","doi-asserted-by":"crossref","first-page":"1275","DOI":"10.1099\/vir.0.18785-0","article-title":"Time-course induction of apoptosis by wild-type and attenuated strains of rubella virus","volume":"84","author":"Cooray","year":"2003","journal-title":"J Gen Virol"},{"key":"R10","doi-asserted-by":"crossref","first-page":"8590","DOI":"10.1038\/sj.onc.1207102","article-title":"The Bcl-2 family: roles in cell survival and oncogenesis","volume":"22","author":"Cory","year":"2003","journal-title":"Oncogene"},{"key":"R11","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1006\/bbrc.1993.2438","article-title":"A new method for cytofluorometric analysis of mitochondrial membrane potential using the J-aggregate forming lipophilic cation 5,5\u2032,6,6'-tetrachloro-1,1',3,3\u2032-tetraethylbenzimidazolcarbocyanineiodide (JC-1)","volume":"197","author":"Cossarizza","year":"1993","journal-title":"Biochem Biophys Res Commun"},{"key":"R12","doi-asserted-by":"crossref","first-page":"W615","DOI":"10.1093\/nar\/gkh398","article-title":"MOLPROBITY: structure validation and all-atom contact analysis for nucleic acids and their complexes","volume":"32","author":"Davis","year":"2004","journal-title":"Nucleic Acids Res"},{"key":"R13","doi-asserted-by":"crossref","first-page":"4738","DOI":"10.1074\/jbc.M411434200","article-title":"Solution structure of prosurvival Mcl-1 and characterization of its binding by proapoptotic BH3-only ligands","volume":"280","author":"Day","year":"2005","journal-title":"J Biol Chem"},{"key":"R14","doi-asserted-by":"crossref","first-page":"36570","DOI":"10.1074\/jbc.M400567200","article-title":"Poxvirus protein N1L targets the I- \u03ba B kinase complex, inhibits signaling to NF- \u03ba B by the tumor necrosis factor superfamily of receptors, and inhibits NF- \u03ba B and IRF3 signaling by toll-like receptors","volume":"279","author":"DiPerna","year":"2004","journal-title":"J Biol Chem"},{"key":"R15","doi-asserted-by":"crossref","first-page":"2126","DOI":"10.1107\/S0907444904019158","article-title":"Coot: model-building tools for molecular graphics","volume":"60","author":"Emsley","year":"2004","journal-title":"Acta Crystallogr D Biol Crystallogr"},{"key":"R16","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1038\/sj.cdd.4401718","article-title":"Modified vaccinia virus Ankara protein F1L is a novel BH3-domain-binding protein and acts together with the early viral protein E3L to block virus-associated apoptosis","volume":"13","author":"Fischer","year":"2006","journal-title":"Cell Death Differ"},{"key":"R17","doi-asserted-by":"crossref","first-page":"5723","DOI":"10.1073\/pnas.97.11.5723","article-title":"Bcl-2 decreases the free Ca2+ concentration within the endoplasmic reticulum","volume":"97","author":"Foyouzi-Youssefi","year":"2000","journal-title":"Proc Natl Acad Sci U S A"},{"key":"R18","doi-asserted-by":"crossref","first-page":"S11","DOI":"10.1017\/S0031182005008127","article-title":"Evasion of innate immunity by vaccinia virus","volume":"130, (Suppl.)","author":"Haga","year":"2005","journal-title":"Parasitology"},{"key":"R19","first-page":"2808","article-title":"Ultrastructural localization of BHRF1: an Epstein-Barr virus gene product which has homology with bcl-2","volume":"54","author":"Hickish","year":"1994","journal-title":"Cancer Res"},{"key":"R20","doi-asserted-by":"crossref","first-page":"1497","DOI":"10.1093\/emboj\/cdg144","article-title":"The structure of Bcl-w reveals a role for the C-terminal residues in modulating biological activity","volume":"22","author":"Hinds","year":"2003","journal-title":"EMBO J"},{"key":"R21","doi-asserted-by":"crossref","first-page":"3428","DOI":"10.1073\/pnas.062525799","article-title":"Solution structure of a Bcl-2 homolog from Kaposi sarcoma virus","volume":"99","author":"Huang","year":"2002","journal-title":"Proc Natl Acad Sci U S A"},{"key":"R22","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1016\/j.jmb.2003.08.007","article-title":"Solution structure of the BHRF1 protein from Epstein-Barr virus, a homolog of human Bcl-2","volume":"332","author":"Huang","year":"2003","journal-title":"J Mol Biol"},{"key":"R23","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1016\/0042-6822(89)90627-2","article-title":"Mapping and insertional mutagenesis of a vaccinia virus gene encoding a 13,800-Da secreted protein","volume":"171","author":"Kotwal","year":"1989","journal-title":"Virology"},{"key":"R24","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/0022-2836(82)90515-0","article-title":"A simple method for displaying the hydropathic character of a protein","volume":"157","author":"Kyte","year":"1982","journal-title":"J Mol Biol"},{"key":"R25","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1016\/S1074-7613(03)00234-6","article-title":"The structure of a Bcl-xL\/Bim fragment complex: implications for Bim function","volume":"19","author":"Liu","year":"2003","journal-title":"Immunity"},{"key":"R26","doi-asserted-by":"crossref","first-page":"e10","DOI":"10.1371\/journal.ppat.0010010","article-title":"A surface groove essential for viral bcl-2 function during chronic infection in vivo","volume":"1","author":"Loh","year":"2005","journal-title":"PLoS Pathog"},{"key":"R27","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1038\/35048069","article-title":"Breaking the mitochondrial barrier","volume":"2","author":"Martinou","year":"2001","journal-title":"Nat Rev Mol Cell Biol"},{"key":"R28","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1038\/381335a0","article-title":"X-ray and NMR structure of Bcl-xL, an inhibitor of programmed cell death","volume":"381","author":"Muchmore","year":"1996","journal-title":"Nature"},{"key":"R29","doi-asserted-by":"crossref","first-page":"4391","DOI":"10.1128\/JVI.67.7.4391-4394.1993","article-title":"An African swine fever virus gene with similarity to the proto-oncogene bcl-2 and the Epstein-Barr virus gene BHRF1","volume":"67","author":"Neilan","year":"1993","journal-title":"J Virol"},{"key":"R30","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1016\/j.jmb.2005.11.032","article-title":"BCL-XL dimerization by three-dimensional domain swapping","volume":"356","author":"O'Neill","year":"2006","journal-title":"J Mol Biol"},{"key":"R31","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1016\/S0076-6879(97)76066-X","article-title":"Processing of X-ray defraction data collected in oscillation mode","volume":"276","author":"Otwinowski","year":"1997","journal-title":"Methods Enzymol"},{"key":"R32","doi-asserted-by":"crossref","first-page":"2528","DOI":"10.1110\/ps.9.12.2528","article-title":"Rationale for Bcl-xL\/Bad peptide complex formation from structure, mutagenesis, and biophysical studies","volume":"9","author":"Petros","year":"2000","journal-title":"Protein Sci"},{"key":"R33","doi-asserted-by":"crossref","first-page":"3012","DOI":"10.1073\/pnas.041619798","article-title":"Solution structure of the antiapoptotic protein bcl-2","volume":"98","author":"Petros","year":"2001","journal-title":"Proc Natl Acad Sci U S A"},{"key":"R34","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.bbamcr.2003.08.012","article-title":"Structural biology of the Bcl-2 family of proteins","author":"Petros","year":"2004","journal-title":"Biochim Biophys Acta"},{"key":"R35","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/j.bbamcr.2003.11.001","article-title":"Viral Bcl-2 homologs and their role in virus replication and associated diseases","author":"Polster","year":"2004","journal-title":"Biochim Biophys Acta"},{"key":"R36","doi-asserted-by":"crossref","first-page":"1651","DOI":"10.1038\/sj.cdd.4401853","article-title":"Interaction of F1L with the BH3 domain of Bak is responsible for inhibiting vaccinia-induced apoptosis","volume":"13","author":"Postigo","year":"2006","journal-title":"Cell Death Differ"},{"key":"R37","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1146\/annurev.micro.53.1.577","article-title":"Viruses and apoptosis","volume":"53","author":"Roulston","year":"1999","journal-title":"Annu Rev Microbiol"},{"key":"R38","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1038\/nm0397-293","article-title":"Kaposi's sarcoma-associated herpesvirus encodes a functional bcl-2 homologue","volume":"3","author":"Sarid","year":"1997","journal-title":"Nat Med"},{"key":"R39","doi-asserted-by":"crossref","first-page":"983","DOI":"10.1126\/science.275.5302.983","article-title":"Structure of Bcl-xL-Bak peptide complex: recognition between regulators of apoptosis","volume":"275","author":"Sattler","year":"1997","journal-title":"Science"},{"key":"R40","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1146\/annurev.immunol.21.120601.141049","article-title":"Poxviruses and immune evasion","volume":"21","author":"Seet","year":"2003","journal-title":"Annu Rev Immunol"},{"key":"R41","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1016\/j.cell.2005.08.012","article-title":"Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF- \u03ba B and IRF3","volume":"122","author":"Seth","year":"2005","journal-title":"Cell"},{"key":"R42","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/0022-2836(79)90416-9","article-title":"Crystal structure of cat muscle pyruvate kinase at a resolution of 2.\u200a\u00c5","volume":"134","author":"Stuart","year":"1979","journal-title":"J Mol Biol"},{"key":"R43","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1016\/S0092-8674(00)00167-7","article-title":"Structure of Bax: coregulation of dimer formation and intracellular localization","volume":"103","author":"Suzuki","year":"2000","journal-title":"Cell"},{"key":"R44","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.virol.2005.09.032","article-title":"Near death experiences: poxvirus regulation of apoptotic death","volume":"344","author":"Taylor","year":"2006","journal-title":"Virology"},{"key":"R45","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1107\/S0907444900005072","article-title":"Maximum-likelihood density modification","volume":"56","author":"Terwilliger","year":"2000","journal-title":"Acta Crystallogr D Biol Crystallogr"},{"key":"R46","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1128\/JVI.78.1.353-366.2004","article-title":"The genome of canarypox virus","volume":"78","author":"Tulman","year":"2004","journal-title":"J Virol"},{"key":"R47","doi-asserted-by":"crossref","first-page":"5894","DOI":"10.1128\/JVI.71.8.5894-5904.1997","article-title":"Complete sequence and genomic analysis of murine gammaherpesvirus 68","volume":"71","author":"Virgin","year":"1997","journal-title":"J Virol"},{"key":"R48","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1107\/S0021889803001997","article-title":"A procedure for setting up high-throughput nanolitre crystallization experiments. I. Protocol design and validation","volume":"36","author":"Walter","year":"2003","journal-title":"J Appl Crystallogr"},{"key":"R49","doi-asserted-by":"crossref","first-page":"14031","DOI":"10.1128\/JVI.79.22.14031-14043.2005","article-title":"The vaccinia virus F1L protein interacts with the proapoptotic protein Bak and inhibits Bak activation","volume":"79","author":"Wasilenko","year":"2005","journal-title":"J Virol"}],"container-title":["Journal of General Virology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jgv\/10.1099\/vir.0.82772-0?crawler=true","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,10,26]],"date-time":"2020-10-26T22:23:48Z","timestamp":1603751028000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jgv\/10.1099\/vir.0.82772-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2007,6,1]]},"references-count":49,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2007,6,1]]}},"URL":"https:\/\/doi.org\/10.1099\/vir.0.82772-0","relation":{},"ISSN":["0022-1317","1465-2099"],"issn-type":[{"value":"0022-1317","type":"print"},{"value":"1465-2099","type":"electronic"}],"subject":[],"published":{"date-parts":[[2007,6,1]]}}}