Escherichia coli replication termination protein impedes the action of helicases.
Identification of the consensus sequence for termination of replication (ter) in Escherichia coli and the isolation of the ter-binding protein (TBP) allowed us to test their effects on replication forks initiated at the unique origin of the E. coli chromosome (oriC) in a purified enzyme system. Replication was severely impeded by ter in a unique orientation when purified TBP was supplied to bind it. The target for blockage within the replication complex can now be ascribed to the inability of dnaB helicase to separate the duplex strands when it encounters ter bound by TBP. Other helicases, such as rep and uvrD proteins, that translocate on DNA and displace strands in the direction opposite to that of dnaB protein are also blocked, but only when the TBP-bound ter is oriented in the other direction. From these results, we infer that the orientation of ter confers a particular polarity on the TBP seated on it, such that a helicase is blocked when it confronts TBP from one side, but can act, presumably by displacing TBP, when facing its other side. Thus, the intrinsic nature of the oriented TBP-ter complex is responsible for impeding the helicases, rather than any protein-protein interactions.
Lee
E H
EH
Department of Biochemistry, Stanford University, CA 94305-5307.
Kornberg
A
A
Hidaka
M
M
Kobayashi
T
T
Horiuchi
T
T
D001483:Base Sequence; D002876:Chromosomes, Bacterial / Q000502:physiology; D004265:DNA Helicases / Q000378:metabolism*; D004261:DNA Replication; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D010957:Plasmids; D015183:Restriction Mapping; D054334:Viral Regulatory and Accessory Proteins / Q000302:isolation & purification / Q000378:metabolism*
D054334:Viral Regulatory and Accessory Proteins; D004265:DNA Helicases