title: Amino acid 55 plays a central role in tetramerization and function of Escherichia coli single-stranded DNA binding protein.
abstract: The histidine at position 55 of the amino acid sequence of the Escherichia coli single-stranded DNA binding protein was replaced by tyrosine, glutamic acid, lysine, phenylalanine, and isoleucine. The properties of the mutant proteins were determined using analytical ultracentrifugation, NMR spectroscopy, gel filtration, and fluorimetric detection of their single-stranded DNA binding ability. While the phenylalanine and isoleucine substitutions did not change the properties of the protein measurably, tyrosine and lysine mutants dissociate into subunits and loose some of their binding affinity for poly(dT). For the lysine mutant we show by electron microscopy that the protein, although fully dissociated and possibly denatured in the free state, binds to poly(dT) as a tetramer indistinguishable from the wild-type protein. The process of tetramerization as observed via single-stranded DNA binding ability is composed of a variety of steps ranging in time from some milliseconds to several hours; it probably involves several forms of dissociated and non-native protein.
authors.affiliation: Medizinische Hochschule, Zentrum Biochemie, Hannover, Federal Republic of Germany.
authors.forename: U
authors.initials: U
authors.lastname: Curth
authors.forename: I
authors.initials: I
authors.lastname: Bayer
authors.forename: J
authors.initials: J
authors.lastname: Greipel
authors.forename: F
authors.initials: F
authors.lastname: Mayer
authors.forename: C
authors.initials: C
authors.lastname: Urbanke
authors.forename: G
authors.initials: G
authors.lastname: Maass
mesh_terms: D004277:DNA, Single-Stranded / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics; D006639:Histidine; D009682:Magnetic Resonance Spectroscopy; D016296:Mutagenesis; D013329:Structure-Activity Relationship
chemical_list: D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D006639:Histidine