A replication terminus located at or near a replication checkpoint of Bacillus subtilis functions independently of stringent control.
We have examined a replication terminus (psiL1) located on the left arm of the chromosome of Bacillus subtilis and within the yxcC gene and at or near the left replication checkpoint that is activated under stringent conditions. The psiL1 sequence appears to bind to two dimers of the replication terminator protein (RTP) rather weakly and seems to possess overlapping core and auxiliary sites that have some sequence similarities with normal Ter sites. Surprisingly, the asymmetrical, isolated psiL1 site arrested replication forks in vivo in both orientations and independent of stringent control. In vitro, the sequence arrested DnaB helicase in both orientations, albeit more weakly than the normal Ter1 terminus. The key points of mechanistic interest that emerge from the present work are: (i) strong binding of a Ter (psiL1) sequence to RTP did not appear to be essential for fork arrest and (ii) polarity of fork arrest could not be correlated in this case with just symmetrical protein-DNA interaction at the core and auxiliary sites of psiL1. On the basis of the result it would appear that the weak RTP-L1Ter interaction cannot by itself account for fork arrest, thus suggesting a role for DnaB-RTP interaction.
Gautam
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Department of Microbiology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Bastia
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D001412:Bacillus subtilis / Q000235:genetics*; D001426:Bacterial Proteins; D001483:Base Sequence; D002876:Chromosomes, Bacterial; D004265:DNA Helicases / Q000037:antagonists & inhibitors; D017931:DNA Primers; D004261:DNA Replication; D004269:DNA, Bacterial / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D053488:DnaB Helicases; D011485:Protein Binding
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C063112:rtP protein, Bacillus subtilis; D004265:DNA Helicases; D053488:DnaB Helicases