title: Oligomerization and DNA-binding capacity of Pmr, a histone-like protein H1 (H-NS) family protein encoded on IncP-7 carbazole-degradative plasmid pCAR1.
abstract: Pmr, a histone-like protein H1 (H-NS) family protein encoded on plasmid pCAR1, is a key factor in optimizing gene transcription on both pCAR1 and the host chromosome. To clarify the mode of function of Pmr, we performed gel filtration chromatography analysis and protein-protein cross-linking, and found that Pmr forms homo-oligomers, consisting of its homodimers. We also found, by atomic force microscopy, that Pmr has DNA-bridging capacity. From these results, Pmr was deduced to have features common to H-NS family proteins. Additionally, evaluating protein-DNA affinity is important to clarify the mode of function of Pmr, and hence we performed an electrophoretic mobility shift assay. Though Pmr formed high-order protein-DNA complexes and did not show preference for nucleic acid sequences, the C-terminal region of Pmr did, suggesting that the DNA-binding affinity of Pmr can be evaluated by using its C-terminal region.
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Japan.
authors.forename: Chiho
authors.initials: C
authors.lastname: Suzuki
authors.forename: Choong-Soo
authors.initials: CS
authors.lastname: Yun
authors.forename: Takashi
authors.initials: T
authors.lastname: Umeda
authors.forename: Tsuguno
authors.initials: T
authors.lastname: Terabayashi
authors.forename: Kazuya
authors.initials: K
authors.lastname: Watanabe
authors.forename: Hisakazu
authors.initials: H
authors.lastname: Yamane
authors.forename: Hideaki
authors.initials: H
authors.lastname: Nojiri
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002227:Carbazoles / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D011550:Pseudomonas aeruginosa; D016958:Pseudomonas putida; D015534:Trans-Activators / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D002227:Carbazoles; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C111692:MvaT protein, Pseudomonas; D015534:Trans-Activators