Role of the amino-terminal domain of bacteriophage phi 29 connector in DNA binding and packaging.
The connector of bacteriophage phi 29 is required for prohead assembly, binds DNA, and drives DNA packaging into viral proheads. Limited proteolysis of the connector protein with endoproteinase Glu-C from Staphylococcus aureus V8 and chymotrypsin showed that a domain of the NH2-terminal region is involved in DNA binding and in the subsequent packaging into preformed proheads, but not in prohead assembly. Mutants in specific amino acids of the NH2-terminal domain, obtained by directed mutagenesis techniques, showed that the Ala1-Arg2-Lys3-Arg4 region of the connector is absolutely necessary for DNA packaging into the proheads as well as for efficient DNA binding.
Donate
L E
LE
Centro de Biología Molecular, Universidad Autónoma de Madrid, Spain.
Valpuesta
J M
JM
Rocher
A
A
Méndez
E
E
Rojo
F
F
Salas
M
M
Carrascosa
J L
JL
D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000235:genetics*; D002213:Capsid / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism*; D005814:Genes, Viral; D006868:Hydrolysis; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D012697:Serine Endopeptidases / Q000378:metabolism
D004279:DNA, Viral; D012697:Serine Endopeptidases; D002918:Chymotrypsin; C028344:glutamyl endopeptidase