title: Conformational changes in bacteriophage phi 29 connector prevents DNA-binding activity.
abstract: In vitro DNA packaging activity in a defined system derived from bacteriophage phi 29 depends upon the chemical integrity of the connector protein p10. Proteolytic cleavage of p10 rendered the proheads inactive for DNA packaging. A similar treatment on isolated connectors abolished the DNA-binding activity of the native p10, but the general shape and size of the connector was not changed as revealed by electron microscopy. Analytical ultracentrifugation showed that the proteolyzed connectors had a smaller sedimentation coefficient, while amino acid analysis after dialysis of the proteolyzed p10 confirmed the loss of 16 and 19 amino acids from the amino and carboxy termini, respectively. Low angle X-ray scattering revealed that proteolysis was followed by a small decrease in the radius of gyration and a reorganization of the distal domain of the cylindrical inner part of the connector. Characterization of the cleavage sites in the primary sequence allowed us to propose the location of the DNA-binding domain in the connector model.
authors.affiliation: Centro de Biología Molecular (CSIC-U AM) Universidad Autónoma de Madrid, Spain.
authors.forename: L
authors.initials: L
authors.lastname: Herranz
authors.forename: J
authors.initials: J
authors.lastname: Bordas
authors.forename: E
authors.initials: E
authors.lastname: Towns-Andrews
authors.forename: E
authors.initials: E
authors.lastname: Mendez
authors.forename: P
authors.initials: P
authors.lastname: Usobiaga
authors.forename: J L
authors.initials: JL
authors.lastname: Carrascosa
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001435:Bacteriophages / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000032:analysis / Q000378:metabolism*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012542:Scattering, Radiation; D014357:Trypsin; D014461:Ultracentrifugation; D014764:Viral Proteins / Q000032:analysis / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D014764:Viral Proteins; D014357:Trypsin