Analysis of interactions among factors involved in the bacteriophage T3 DNA packaging reaction in a defined in vitro system.
During head assembly of phage T3, DNA is packaged into the cavity of a preformed protein shell, called the prohead, with the aid of noncapsid, packaging proteins, the products of genes 18 and 19 (gp18 and gp19). gp18 and gp19 separately form complexes with DNA and proheads, respectively. These complexes associate to form a precursor which can be converted to filled heads by the addition of ATP. Interactions among factors involved in DNA packaging were analyzed. In the presence of ATP, gp19 formed functional complexes with proheads. Formation of gp19-prohead complex showed a sigmoidal dependence on ATP concentration with a half maximal concentration of about 7.5 microM. Six molecules of gp19 bound to the prohead at a saturating amount of gp19. gp19 did not bind to proheads lacking the connector of gp8 (8- prohead). In the absence of ATP, proheads were inactivated by gp19. The gp19-prohead complexes formed in the absence of ATP contained 20-30 gp19 molecules per prohead and formed multimeric aggregates. 8- proheads did not bind gp19 and did not form such aggregates even in the absence of ATP. From these results, we conclude that 6 molecules of gp19 bind to the gp8 connector structure in the portal vertex of the prohead. The cleavage patterns of gp19 by several proteases were altered by the addition of ATP, indicating that ATP induces a conformational change in gp19, gp18 bound only to linear, duplex DNA.
Fujisawa
H
H
Department of Botany, Faculty of Science, Kyoto University, Japan.
Shibata
H
H
Kato
H
H
D000255:Adenosine Triphosphate / Q000494:pharmacology; D002918:Chymotrypsin / Q000187:drug effects / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure; D007700:Kinetics; D008854:Microscopy, Electron; D009690:Nucleic Acid Conformation; D013604:T-Phages / Q000235:genetics / Q000378:metabolism*; D014357:Trypsin / Q000187:drug effects / Q000378:metabolism; D014764:Viral Proteins / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure
D004279:DNA, Viral; D004268:DNA-Binding Proteins; D014764:Viral Proteins; D000255:Adenosine Triphosphate; D002918:Chymotrypsin; D014357:Trypsin