The degree of oligomerization of the H-NS nucleoid structuring protein is related to specific binding to DNA
| Autor: | Roy Williams, Sylvie Rimsky, Emeline Bouffartigues, Bruno Robert, Véronique Arluison, Henri Buc, Cyril Badaut |
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| Přispěvatelé: | Laboratoire de Biologie et de Pharmacologie Appliquée ( LBPA ), École normale supérieure - Cachan ( ENS Cachan ) -Centre National de la Recherche Scientifique ( CNRS ), Physicochimie des Macromolecules Biologiques, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Laboratoire Bioénergétique, Métalloprotéines et Stress ( LBMS ), Département Biochimie, Biophysique et Biologie Structurale ( B3S ), Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Sud - Paris 11 ( UP11 ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Université Paris-Sud - Paris 11 ( UP11 ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Sud - Paris 11 ( UP11 ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Université Paris-Sud - Paris 11 ( UP11 ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Biologie et de Pharmacologie Appliquée (LBPA), Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Cachan (ENS Cachan), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Laboratoire Bioénergétique Membranaire et Stress (LBMS), Département Biochimie, Biophysique et Biologie Structurale (B3S), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), École normale supérieure - Cachan (ENS Cachan)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2002 |
| Předmět: |
MESH: Amino Acid Transport Systems
Amino Acid Transport Systems MESH : DNA Mutant MESH : Amino Acid Transport Systems Electrophoretic Mobility Shift Assay Biochemistry chemistry.chemical_compound Transcription (biology) MESH : Bacterial Proteins Promoter Regions Genetic MESH: Bacterial Proteins MESH : Temperature Coiled coil 0303 health sciences MESH: DNA Temperature MESH: Temperature DNA-Binding Proteins MESH: Promoter Regions (Genetics) Cross-Linking Reagents MESH : Dimerization MESH : Electrophoretic Mobility Shift Assay MESH : Deoxyribonuclease I MESH : DNA-Binding Proteins Dimerization MESH : Cross-Linking Reagents H-NS MESH: Cross-Linking Reagents Biology MESH : Ultracentrifugation bacterial chromatine 03 medical and health sciences Bacterial Proteins Nucleoid Deoxyribonuclease I [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Molecular Biology [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry Molecular Biology 030304 developmental biology 030306 microbiology DNase-I Footprinting MESH: Ultracentrifugation Promoter Cell Biology DNA MESH: Deoxyribonuclease I MESH : Promoter Regions (Genetics) Nucleoprotein Crystallography chemistry MESH: Dimerization MESH: Electrophoretic Mobility Shift Assay Biophysics Ultracentrifugation MESH: DNA-Binding Proteins |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2002, 277 (44), pp.41657-66. 〈10.1074/jbc.M206037200〉 Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2002, 277 (44), pp.41657-66. ⟨10.1074/jbc.M206037200⟩ Journal of Biological Chemistry, 2002, 277 (44), pp.41657-66. ⟨10.1074/jbc.M206037200⟩ |
| ISSN: | 0021-9258 1083-351X |
| Popis: | International audience; At several E. coli promoters, initiation of transcription is repressed by a tight nucleoprotein complex formed by the assembly of the H-NS protein. In order to characterize the relationship between the structure of H-NS oligomers in solution and on relevant DNA fragments, we have compared wild-type H-NS and several transdominant H-NS mutants using gel shift assays, DNase I footprinting, analytical ultracentrifugation, and reactivity toward a cross-linking reagent. In solution, oligomerization occurs through two protein interfaces, one necessary to construct a dimeric core (and involving residues 1-64) and the other required for subsequent assembly of these dimers. We show that, as well as region 64-95, residues present in the NH(2)-terminal coiled coil domain also participate in this second interface. Our results support the view that the same interacting interfaces are also involved on the DNA. We propose that the dimeric core recognizes specific motifs, with the second interface being critical for their correct head to tail assembly. The COOH-terminal domain of the protein contains the DNA binding motif essential for the discrimination of this specific functional assembly over competitive nonspecific H-NS polymers. |
| Databáze: | OpenAIRE |
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