Two Polymorphic Residues Account for the Differences in DNA Binding and Transcriptional Activation by NF-κB Proteins Encoded by Naturally Occurring Alleles in Nematostella vectensis
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The NF-κB family of transcription factors is activated in response to many environmental and biological stresses, and plays a key role in innate immunity across a broad evolutionary expanse of animals. A simple NF-κB pathway is present in the sea anemone Nematostella vectensis, an important model organism in the phylum Cnidaria. Nematostella has previously been shown to have two naturally occurring NF-κB alleles (Nv-NF-κB-C and Nv-NF-κB-S) that encode proteins with different DNA-binding and transactivation abilities. We show here that polymorphic residues 67 (Cys vs. Ser) and 269 (Ala vs. Glu) play complementary roles in determining the DNA-binding activity of the NF-κB proteins encoded by these two alleles and that residue 67 is primarily responsible for the difference in their transactivation ability. Phylogenetic analysis indicates that Nv-NF-κB-S is the derived allele, consistent with its restricted geographic distribution. These results define polymorphic residues that are important for the DNA-binding and transactivating activities of two naturally occurring variants of Nv-NF-κB. The implications for the appearance of the two Nv-NF-κB alleles in natural populations of sea anemones are discussed.
KeywordsNF-κB Nematostella Polymorphism Evolution DNA binding Transactivation
This research was supported by grant MCB-0920461 from the National Science Foundation (J.R.F., T.D.G.) and ARRA supplement CA047763-22S3 (to T.D.G.). F.S.W. was supported by predoctoral grant by the Superfund Basic Research Program at Boston University 5 P42 ES07381, and F.S.W. and D.J.S. were supported by Warren-McLeod Fellowships. N.J. was supported by funds from the Boston University Undergraduate Research Opportunities Program. We thank Tristan Lubinski and Lauren Friedman for help with bioinformatic analyses and helpful discussions.
- Abramoff MD, Magelhaes PJ, Ram SJ (2004) Image processing with ImageJ. J Biophotonics 11(7):36–42Google Scholar
- Brooks BR, Brooks CL III, Mackerell AD Jr, Nilsson L, Petrella RJ, Roux B, Won Y, Archontis G, Bartels C, Boresch S, Caflisch A, Caves L, Cui Q, Dinner AR, Feig M, Fischer S, Gao J, Hodoscek M, Im W, Kuczera K, Lazaridis T, Ma J, Ovchinnikov V, Paci E, Pastor RW, Post CB, Pu JZ, Schaefer M, Tidor B, Venable RM, Woodcock HL, Wu X, Yang W, York DM, Karplus M (2009) CHARMM: the biomolecular simulation program. J Comput Chem 30(10):1545–1614. doi: 10.1002/jcc.21287 PubMedCrossRefGoogle Scholar
- Ertürk-Hasdemir D, Broemer M, Leulier F, Lane WS, Paquette N, Hwang D, Kim CH, Stoven S, Meier P, Silverman N (2009) Two roles for the Drosophila IKK complex in the activation of Relish and the induction of antimicrobial peptide genes. Proc Natl Acad Sci USA 106(24):9779–9784. doi: 10.1073/pnas.0812022106 PubMedCrossRefGoogle Scholar
- Gilmore TD, Wolenski FS (2012) NF-κB: where did it come from and why? Immunol Rev (in press)Google Scholar
- Liang M-C, Bardhan S, Pace EA, Rosman D, Beutler JA, Porco JA Jr, Gilmore TD (2006a) Inhibition of transcription factor NF-κB signaling proteins IKKβ and p65 through specific cysteine residues by epoxyquinone A monomer: correlation with its anti-cancer cell growth activity. Biochem Pharmacol 71(5):634–645. doi: 10.1016/j.bcp.2005.11.013 PubMedCrossRefGoogle Scholar
- Liang M-C, Bardhan S, Porco JA Jr, Gilmore TD (2006b) The synthetic epoxyquinoids jesterone dimer and epoxyquinone A monomer induce apoptosis and inhibit REL (human c-Rel) DNA binding in an IκBα-deficient diffuse large B-cell lymphoma cell line. Cancer Lett 241(1):69–78. doi: 10.1016/j.canlet.2005.10.004 PubMedCrossRefGoogle Scholar
- Shinzato C, Shoguchi E, Kawashima T, Hamada M, Hisata K, Tanaka M, Fujie M, Fujiwara M, Koyanagi R, Ikuta T, Fujiyama A, Miller DJ, Satoh N (2011) Using the Acropora digitifera genome to understand coral responses to environmental change. Nature 476(7360):320–323. doi: 10.1038/nature10249 PubMedCrossRefGoogle Scholar
- Sullivan JC, Wolenski FS, Reitzel AM, French CE, Traylor-Knowles N, Gilmore TD, Finnerty JR (2009) Two alleles of NF-κB in the sea anemone Nematostella vectensis are widely dispersed in nature and encode proteins with distinct activities. PLoS ONE 4(10):e7311. doi: 10.1371/journal.pone.0007311 PubMedCrossRefGoogle Scholar
- Torruella G, Derelle R, Paps J, Lang BF, Roger AJ, Shalchian-Tabrizi K, Ruiz-Trillo I (2011) Phylogenetic relationships within the Opisthokonta based on phylogenomic analyses of conserved single copy protein domains. Mol Biol Evol. doi: 10.1093/molbev/msr185
- Wolenski FS, Garbati MR, Lubinski TJ, Traylor-Knowles N, Dresselhaus E, Stefanik DJ, Goucher H, Finnerty JR, Gilmore TD (2011) Characterization of the core elements of the NF-κB signaling pathway of the sea anemone Nematostella vectensis. Mol Cell Biol 31(5):1076–1087. doi: 10.1128/mcb.00927-10 PubMedCrossRefGoogle Scholar