The functional asymmetry of cosN, the nicking site for bacteriophage lambda DNA packaging, is dependent on the terminase binding site, cosB.
cosN is the site at which terminase, the DNA packaging enzyme of phage lambda, introduces staggered nicks into viral concatemeric DNA to initiate genome packaging. Although the nick positions and many of the base pairs of cosN show 2-fold rotational symmetry, cosN is functionally asymmetric. That is, the cosN G2C mutation in the left half-site (cosNL) causes a strong virus growth defect whereas the symmetrically disposed cosN C11G mutation in the right half-site (cosNR) does not affect virus growth. The experiments reported here test the proposal that the genetic asymmetry of cosN results from terminase interactions with cosB, a binding site to the right of cosN. In the presence of cosB, the left half-site mutation, cosN G2C, strongly affected the cos cleavage reaction, while the symmetric right half-site mutation, cosN C11G, had little effect. In the absence of cosB, the two mutations moderately reduced the rate of cos cleavage by the same amount. The results indicated that the functional asymmetry of cosNdepends on the presence of cosB. A model is discussed in which terminase-cosN interactions in the nicking complex are assisted by anchoring of terminase to cosB.
Hang
J Q
JQ
Molecular Biology Program and Department of Microbiology, University of Iowa, Iowa City, Iowa 52242, USA.
Catalano
C E
CE
Feiss
M
M
D010582:Bacteriophage lambda / Q000235:genetics*; D001665:Binding Sites; D017931:DNA Primers / Q000737:chemistry; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000235:genetics*; D004706:Endodeoxyribonucleases / Q000235:genetics*; D007700:Kinetics; D008958:Models, Molecular; D010957:Plasmids; D011485:Protein Binding; D021122:Protein Subunits; D014764:Viral Proteins / Q000235:genetics*; D014779:Virus Replication
D017931:DNA Primers; D004274:DNA, Recombinant; D004279:DNA, Viral; D021122:Protein Subunits; D014764:Viral Proteins; D004706:Endodeoxyribonucleases