Identification of a compact DNA-binding domain in the gene 5 protein of Pf1 bacteriophage.
The structure of the gene 5 protein of filamentous bacteriophage Pf1 and its interaction with viral DNA have been investigated by a series of limited proteolysis experiments. The ability of purified proteolytic fragments of the Pf1 gene 5 protein to bind oligonucleotides and polynucleotides was monitored by gel retardation and fluorescence. The results show the presence of a compact DNA-binding "core" domain consisting of residues 1-112 of the protein, which is protected from proteolysis in the nucleoprotein complex. Digestion of the free gene 5 protein with subtilisin produces a smaller fragment (residues 7-102) which can no longer bind DNA. Although the N-terminal "core" domain shows full DNA binding activity by fluorescence, the gel retardation experiments suggest reduced kinetic stability of this domain in complexes with oligonucleotides, resulting from the removal of residues 113-144 from the C-terminus of the protein. The sequence of the C-terminal 32 amino acid residues is unusual, with a high proportion of alanine, glutamine, and proline residues which may be related to the role of this sequence in stabilizing the complex.
Plyte
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Biophysics Laboratory, School of Biological Sciences, University of Portsmouth, U.K.
Kneale
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D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000235:genetics*; D001665:Binding Sites; D002268:Carboxypeptidases / Q000378:metabolism; D002918:Chymotrypsin / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism*; D010450:Endopeptidases / Q000378:metabolism; D005814:Genes, Viral; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009698:Nucleoproteins / Q000378:metabolism; D010206:Papain / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013381:Subtilisins / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism
D004279:DNA, Viral; D009698:Nucleoproteins; D010446:Peptide Fragments; D014764:Viral Proteins; C069558:gene V protein, Enterobacteria phage f1; D002268:Carboxypeptidases; D010450:Endopeptidases; C022633:serine carboxypeptidase; D013381:Subtilisins; D002918:Chymotrypsin; D014357:Trypsin; D010206:Papain