title: A Phylogenetically Conserved Group of Nuclear Factor-Y Transcription Factors Interact to Control Nodulation in Legumes.
abstract: The endosymbiotic association between legumes and soil bacteria called rhizobia leads to the formation of a new root-derived organ called the nodule in which differentiated bacteria convert atmospheric nitrogen into a form that can be assimilated by the host plant. Successful root infection by rhizobia and nodule organogenesis require the activation of symbiotic genes that are controlled by a set of transcription factors (TFs). We recently identified Medicago truncatula nuclear factor-YA1 (MtNF-YA1) and MtNF-YA2 as two M. truncatula TFs playing a central role during key steps of the Sinorhizobium meliloti-M. truncatula symbiotic interaction. NF-YA TFs interact with NF-YB and NF-YC subunits to regulate target genes containing the CCAAT box consensus sequence. In this study, using a yeast two-hybrid screen approach, we identified the NF-YB and NF-YC subunits able to interact with MtNF-YA1 and MtNF-YA2. In yeast (Saccharomyces cerevisiae) and in planta, we further demonstrated by both coimmunoprecipitation and bimolecular fluorescence complementation that these NF-YA, -B, and -C subunits interact and form a stable NF-Y heterotrimeric complex. Reverse genetic and chromatin immunoprecipitation-PCR approaches revealed the importance of these newly identified NF-YB and NF-YC subunits for rhizobial symbiosis and binding to the promoter of MtERN1 (for Ethylene Responsive factor required for Nodulation), a direct target gene of MtNF-YA1 and MtNF-YA2. Finally, we verified that a similar trimer is formed in planta by the common bean (Phaseolus vulgaris) NF-Y subunits, revealing the existence of evolutionary conserved NF-Y protein complexes to control nodulation in leguminous plants. This sheds light on the process whereby an ancient heterotrimeric TF mainly controlling cell division in animals has acquired specialized functions in plants.
authors.affiliation: Laboratoire des Interactions Plantes-Microorganismes, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2594, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Laboratoire des Interactions Plantes-Microorganismes, Institut National de la Recherche Agronomique, Unité Mixte de Recherche 441, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Económicas, Universidad Nacional de La Plata, CP 1900 La Plata, Argentina (C.R., F.A.B., M.E.Z.); andCentre National de la Recherche Scientifique, Institute of Plant Sciences, Université Paris-Saclay, 91405 Orsay, France (F.A., M.C.).
authors.forename: Maël
authors.identifier: http://orcid.org/0000-0001-8103-3813
authors.initials: M
authors.lastname: Baudin
authors.affiliation: Laboratoire des Interactions Plantes-Microorganismes, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2594, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Laboratoire des Interactions Plantes-Microorganismes, Institut National de la Recherche Agronomique, Unité Mixte de Recherche 441, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Económicas, Universidad Nacional de La Plata, CP 1900 La Plata, Argentina (C.R., F.A.B., M.E.Z.); andCentre National de la Recherche Scientifique, Institute of Plant Sciences, Université Paris-Saclay, 91405 Orsay, France (F.A., M.C.).
authors.forename: Tom
authors.initials: T
authors.lastname: Laloum
authors.affiliation: Laboratoire des Interactions Plantes-Microorganismes, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2594, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Laboratoire des Interactions Plantes-Microorganismes, Institut National de la Recherche Agronomique, Unité Mixte de Recherche 441, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Económicas, Universidad Nacional de La Plata, CP 1900 La Plata, Argentina (C.R., F.A.B., M.E.Z.); andCentre National de la Recherche Scientifique, Institute of Plant Sciences, Université Paris-Saclay, 91405 Orsay, France (F.A., M.C.).
authors.forename: Agnès
authors.initials: A
authors.lastname: Lepage
authors.affiliation: Laboratoire des Interactions Plantes-Microorganismes, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2594, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Laboratoire des Interactions Plantes-Microorganismes, Institut National de la Recherche Agronomique, Unité Mixte de Recherche 441, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Económicas, Universidad Nacional de La Plata, CP 1900 La Plata, Argentina (C.R., F.A.B., M.E.Z.); andCentre National de la Recherche Scientifique, Institute of Plant Sciences, Université Paris-Saclay, 91405 Orsay, France (F.A., M.C.).
authors.forename: Carolina
authors.identifier: http://orcid.org/0000-0001-8492-2667
authors.initials: C
authors.lastname: Rípodas
authors.affiliation: Laboratoire des Interactions Plantes-Microorganismes, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2594, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Laboratoire des Interactions Plantes-Microorganismes, Institut National de la Recherche Agronomique, Unité Mixte de Recherche 441, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Económicas, Universidad Nacional de La Plata, CP 1900 La Plata, Argentina (C.R., F.A.B., M.E.Z.); andCentre National de la Recherche Scientifique, Institute of Plant Sciences, Université Paris-Saclay, 91405 Orsay, France (F.A., M.C.).
authors.forename: Federico
authors.initials: F
authors.lastname: Ariel
authors.affiliation: Laboratoire des Interactions Plantes-Microorganismes, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2594, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Laboratoire des Interactions Plantes-Microorganismes, Institut National de la Recherche Agronomique, Unité Mixte de Recherche 441, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Económicas, Universidad Nacional de La Plata, CP 1900 La Plata, Argentina (C.R., F.A.B., M.E.Z.); andCentre National de la Recherche Scientifique, Institute of Plant Sciences, Université Paris-Saclay, 91405 Orsay, France (F.A., M.C.).
authors.forename: Lisa
authors.initials: L
authors.lastname: Frances
authors.affiliation: Laboratoire des Interactions Plantes-Microorganismes, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2594, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Laboratoire des Interactions Plantes-Microorganismes, Institut National de la Recherche Agronomique, Unité Mixte de Recherche 441, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Económicas, Universidad Nacional de La Plata, CP 1900 La Plata, Argentina (C.R., F.A.B., M.E.Z.); andCentre National de la Recherche Scientifique, Institute of Plant Sciences, Université Paris-Saclay, 91405 Orsay, France (F.A., M.C.).
authors.forename: Martin
authors.identifier: http://orcid.org/0000-0002-0310-2163
authors.initials: M
authors.lastname: Crespi
authors.affiliation: Laboratoire des Interactions Plantes-Microorganismes, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2594, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Laboratoire des Interactions Plantes-Microorganismes, Institut National de la Recherche Agronomique, Unité Mixte de Recherche 441, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Económicas, Universidad Nacional de La Plata, CP 1900 La Plata, Argentina (C.R., F.A.B., M.E.Z.); andCentre National de la Recherche Scientifique, Institute of Plant Sciences, Université Paris-Saclay, 91405 Orsay, France (F.A., M.C.).
authors.forename: Pascal
authors.initials: P
authors.lastname: Gamas
authors.affiliation: Laboratoire des Interactions Plantes-Microorganismes, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2594, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Laboratoire des Interactions Plantes-Microorganismes, Institut National de la Recherche Agronomique, Unité Mixte de Recherche 441, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Económicas, Universidad Nacional de La Plata, CP 1900 La Plata, Argentina (C.R., F.A.B., M.E.Z.); andCentre National de la Recherche Scientifique, Institute of Plant Sciences, Université Paris-Saclay, 91405 Orsay, France (F.A., M.C.).
authors.forename: Flavio Antonio
authors.identifier: http://orcid.org/0000-0002-8380-8472
authors.initials: FA
authors.lastname: Blanco
authors.affiliation: Laboratoire des Interactions Plantes-Microorganismes, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2594, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Laboratoire des Interactions Plantes-Microorganismes, Institut National de la Recherche Agronomique, Unité Mixte de Recherche 441, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Económicas, Universidad Nacional de La Plata, CP 1900 La Plata, Argentina (C.R., F.A.B., M.E.Z.); andCentre National de la Recherche Scientifique, Institute of Plant Sciences, Université Paris-Saclay, 91405 Orsay, France (F.A., M.C.).
authors.forename: Maria Eugenia
authors.identifier: http://orcid.org/0000-0001-9565-1743
authors.initials: ME
authors.lastname: Zanetti
authors.affiliation: Laboratoire des Interactions Plantes-Microorganismes, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2594, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Laboratoire des Interactions Plantes-Microorganismes, Institut National de la Recherche Agronomique, Unité Mixte de Recherche 441, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Económicas, Universidad Nacional de La Plata, CP 1900 La Plata, Argentina (C.R., F.A.B., M.E.Z.); andCentre National de la Recherche Scientifique, Institute of Plant Sciences, Université Paris-Saclay, 91405 Orsay, France (F.A., M.C.).
authors.forename: Fernanda
authors.initials: F
authors.lastname: de Carvalho-Niebel
authors.affiliation: Laboratoire des Interactions Plantes-Microorganismes, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2594, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Laboratoire des Interactions Plantes-Microorganismes, Institut National de la Recherche Agronomique, Unité Mixte de Recherche 441, F-31326 Castanet-Tolosan, France (M.B., T.L., A.L., L.F., P.G., F.d.C.-N., A.N.);Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Económicas, Universidad Nacional de La Plata, CP 1900 La Plata, Argentina (C.R., F.A.B., M.E.Z.); andCentre National de la Recherche Scientifique, Institute of Plant Sciences, Université Paris-Saclay, 91405 Orsay, France (F.A., M.C.) andreas.niebel@toulouse.inra.fr.
authors.forename: Andreas
authors.identifier: http://orcid.org/0000-0002-3402-8381
authors.initials: A
authors.lastname: Niebel
mesh_terms: D000595:Amino Acid Sequence; D023081:CCAAT-Binding Factor / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D003593:Cytoplasm / Q000235:genetics / Q000378:metabolism; D007887:Fabaceae / Q000235:genetics* / Q000378:metabolism / Q000382:microbiology; D020869:Gene Expression Profiling / Q000379:methods; D018506:Gene Expression Regulation, Plant; D054884:Host-Pathogen Interactions; D046913:Medicago truncatula / Q000235:genetics / Q000382:microbiology; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D027805:Phaseolus / Q000235:genetics / Q000382:microbiology; D010802:Phylogeny; D010940:Plant Proteins / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D055170:Plant Root Nodulation / Q000235:genetics*; D011485:Protein Binding; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012231:Rhizobium / Q000502:physiology; D017386:Sequence Homology, Amino Acid; D016962:Sinorhizobium meliloti / Q000502:physiology; D013559:Symbiosis; D014157:Transcription Factors / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D020798:Two-Hybrid System Techniques
chemical_list: D023081:CCAAT-Binding Factor; D010940:Plant Proteins; D014157:Transcription Factors; C402244:nuclear factor Y