TY - JOUR
T1 - Sequence and structure of cmp, the replication enhancer of the Staphylococcus aureus plasmid pT181
AU - Henriquez, Vitalia
AU - Milisavljevic, Vladana
AU - Kahn, Jason D.
AU - Gennaro, Maria Laura
N1 - Funding Information:
We wish to thank S. Adhya for providing plasmid pBend2, and K. Drlica and J. Kornblum for critically reading the manuscript. This work was supported by a research grant from the National Science Foundation to M.L.G. (DMB-9018718). J.D.K. acknowledges support from National Institutes of Health Postdoctoral Fellowship GM14122 and NIH grant GM21966 to D.M. Crothers.
PY - 1993/11/30
Y1 - 1993/11/30
N2 - The Staphylococcus aureus plasmid pT181 possesses a DNA replication enhancer element, called cmp, that is required in cis for optimal utilization of the initiator protein by the origin of replication. The minimal nucleotide sequence required for cmp activity was defined by testing progressively smaller DNA fragments for their ability to restore cmp activity in a plasmid mutant deleted for cmp. These experiments indicate that cmp is a sequence of 100 base pairs (bp) characterized by a loosely repeated sequence motif and phased oligo(dT) tracts. Intrinsic DNA bending at cmp was detected by a circular permutation assay of the locus using polyacrylamide-gel electrophoresis and by computer modeling. The cmp element was found to contain two loci of intrinsically bent DNA that confer an overall bent conformation to this replication enhancer.
AB - The Staphylococcus aureus plasmid pT181 possesses a DNA replication enhancer element, called cmp, that is required in cis for optimal utilization of the initiator protein by the origin of replication. The minimal nucleotide sequence required for cmp activity was defined by testing progressively smaller DNA fragments for their ability to restore cmp activity in a plasmid mutant deleted for cmp. These experiments indicate that cmp is a sequence of 100 base pairs (bp) characterized by a loosely repeated sequence motif and phased oligo(dT) tracts. Intrinsic DNA bending at cmp was detected by a circular permutation assay of the locus using polyacrylamide-gel electrophoresis and by computer modeling. The cmp element was found to contain two loci of intrinsically bent DNA that confer an overall bent conformation to this replication enhancer.
KW - DNA bending
KW - DNA deletion analysis
KW - Replication origin
KW - circular permutation assay
KW - initiator protein
UR - http://www.scopus.com/inward/record.url?scp=0027382431&partnerID=8YFLogxK
U2 - 10.1016/0378-1119(93)90179-7
DO - 10.1016/0378-1119(93)90179-7
M3 - Article
C2 - 8244037
AN - SCOPUS:0027382431
SN - 0378-1119
VL - 134
SP - 93
EP - 98
JO - Gene
JF - Gene
IS - 1
ER -