Distamycin-induced inhibition of formation of a nucleoprotein complex between the terminase small subunit G1P and the non-encapsidated end (pacL site) of Bacillus subtilis bacteriophage SPP1.
The small subunit of the Bacillus subtilis bacteriophage SPP1 terminase (G1P) forms a sequence-specific nucleoprotein complex with the SPP1 non-encapsidated end (pacL site) during initiation of DNA encapsidation. Gel mobility shift assay was used to study the G1P-pacL interaction. Distamycin, a minor groove binder that induces local distortion of the DNA, inhibits G1P-pacL complex formation. The competition of G1P with distamycin for DNA binding at the pacL site is independent of the order of addition of the reactants. Other minor groove binders, such as spermine or Hoechst 33258, which do not distort DNA, failed to compete with G1P for pacL DNA binding. Cationic metals, which generate a repertoire of DNA structures different from that caused by the minor groove binders, can partially reverse the distamycin-induced inhibition of G1P binding to pacL DNA. The major groove binder methyl green, which does not distort sequence-directed bending of pacL DNA, competes with G1P for binding at the pacL site. Our data suggest that the natural sequence-directed bend that exists within the pacL site is the architectural element that facilitates assembly of a nucleoprotein complex and hence initiation of DNA encapsidation by bacteriophage SPP1.
Chai
S
S
Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma de Madrid, Cantoblanco, Madrid, Spain.
Alsonso
J C
JC
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D017103:Bacillus Phages / Q000201:enzymology*; D001412:Bacillus subtilis / Q000821:virology; D006690:Bisbenzimidazole / Q000494:pharmacology; D002413:Cations, Divalent; D004279:DNA, Viral / Q000737:chemistry / Q000378:metabolism*; D004214:Distamycins / Q000494:pharmacology*; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D008739:Methyl Green / Q000494:pharmacology; D009690:Nucleic Acid Conformation / Q000187:drug effects; D009698:Nucleoproteins / Q000187:drug effects*; D011485:Protein Binding / Q000187:drug effects; D012394:Rosaniline Dyes / Q000494:pharmacology; D013096:Spermine / Q000494:pharmacology; D019065:Virus Assembly
D000900:Anti-Bacterial Agents; D002413:Cations, Divalent; D004279:DNA, Viral; D004214:Distamycins; D009698:Nucleoproteins; D012394:Rosaniline Dyes; D013096:Spermine; D008739:Methyl Green; D004706:Endodeoxyribonucleases; C030894:terminase; D006690:Bisbenzimidazole