title: Evolutionarily conserved primary TNF sequences relate to its primitive functions in cell death induction.
abstract: TNF is a primitive protein that has emerged from more than 550 million years of evolution. Our bioinformatics study of TNF from nine different taxa in vertebrates revealed several conserved regions in the TNF sequence. By screening overlapping peptides derived from human TNF to determine their role in three different TNF-induced processes--apoptosis, necrosis and NF-κB stimulation--we found that TNF conserved regions are mostly related to cell death rather than NF-κB stimulation. Among the most conserved regions, peptides (P)12, P13 and P1213 (comprising P12 and P13) induced apoptosis, whereas P14, P15, P16 and P1516 (comprising P15 and P16) induced necrosis. Cell death induced by these peptides was not through binding to the TNF receptor. P16-induced necrosis was mainly through disruption of the cell membrane, whereas P1213-induced apoptosis involved activation of TRADD followed by formation of complex II. Finally, using a monoclonal antibody and a mutant TNF protein, we show that TNF-induced apoptosis is determined by a conserved linear sequence that corresponds to that within P1213. Our results reveal the determinant sequence that is key to the TNF primitive function of inducing apoptosis.
authors.affiliation: Department of Oncology, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK.
authors.forename: Wenshu
authors.initials: W
authors.lastname: Lu
authors.affiliation: Department of Oncology, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK.
authors.forename: Qiongyu
authors.initials: Q
authors.lastname: Chen
authors.affiliation: Department of Oncology, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK.
authors.forename: Songmin
authors.initials: S
authors.lastname: Ying
authors.affiliation: Department of Oncology, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK.
authors.forename: Xiaobing
authors.initials: X
authors.lastname: Xia
authors.affiliation: MRC Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Headington OX3 9DS, UK.
authors.forename: Zhanru
authors.initials: Z
authors.lastname: Yu
authors.affiliation: MRC Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Headington OX3 9DS, UK.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Lui
authors.affiliation: Chiralabs Limited, Begbroke Science Park, Woodstock Road, Begbroke, Oxfordshire OX5 1PF, UK.
authors.forename: George
authors.initials: G
authors.lastname: Tranter
authors.affiliation: Department of Immunology, Fourth Military Medical University, Xi'an City 710032, Shaanxi Province, China.
authors.forename: Boquan
authors.initials: B
authors.lastname: Jin
authors.affiliation: Department of Immunology, Fourth Military Medical University, Xi'an City 710032, Shaanxi Province, China.
authors.forename: Chaojun
authors.initials: C
authors.lastname: Song
authors.affiliation: Department of Oncology, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK len.seymour@oncology.ox.ac.uk shisong.jiang@oncology.ox.ac.uk.
authors.forename: Leonard W
authors.initials: LW
authors.lastname: Seymour
authors.affiliation: Department of Oncology, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK len.seymour@oncology.ox.ac.uk shisong.jiang@oncology.ox.ac.uk.
authors.forename: Shisong
authors.initials: S
authors.lastname: Jiang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D053181:Caspase 8 / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D017124:Conserved Sequence; D019143:Evolution, Molecular; D053200:Fas-Associated Death Domain Protein / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D008969:Molecular Sequence Data; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D016601:RNA-Binding Proteins / Q000378:metabolism; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000378:metabolism; D053199:TNF Receptor-Associated Death Domain Protein / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry* / Q000378:metabolism*; D014714:Vertebrates
chemical_list: C119724:AGFG1 protein, human; C505533:FADD protein, human; D053200:Fas-Associated Death Domain Protein; D028861:Nuclear Pore Complex Proteins; D010455:Peptides; D016601:RNA-Binding Proteins; D047888:Receptors, Tumor Necrosis Factor, Type I; D053199:TNF Receptor-Associated Death Domain Protein; D014409:Tumor Necrosis Factor-alpha; D053181:Caspase 8
keywords: Apoptosis; Evolution; NF-κB; Necrosis; Overlapping peptides; TNF