Two Modes of PriA Binding to DNA

Nurse, Pearl; Liu, Joing; Marians, Kenneth J.

doi:10.1074/jbc.274.35.25026

Cited by 91 publications

(149 citation statements)

References 38 publications

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“…This is consistent with its substrate specificity in binding to D loops, but not bubbles, and three-stranded structures that provide models for replication forks (7,8). PriA contains an intrinsic 3Ј-5Ј helicase that has been suggested to function to clear an annealed lagging strand product from the replication fork, creating a site for primosome assembly and helicase loading (9,10).…”

mentioning

confidence: 57%

The PriA Replication Restart Protein Blocks Replicase Access Prior to Helicase Assembly and Directs Template Specificity through Its ATPase Activity

Manhart

McHenry

2013

Journal of Biological Chemistry

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Background:PriA initiates restart at stalled replication forks. Results: A FRET assay detects E. coli and B. subtilis PriA-mediated helicase loading. Conclusion: PriA ATPase activity directs specificity toward larger gaps on leading strands. PriA blocks replicase binding. Significance: This work reveals a novel role of an ATPase in regulating DNA structural specificity and establishes the mechanism of PriA as a checkpoint protein.

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mentioning

confidence: 57%

The PriA Replication Restart Protein Blocks Replicase Access Prior to Helicase Assembly and Directs Template Specificity through Its ATPase Activity

Manhart

McHenry

2013

Journal of Biological Chemistry

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“…How can replication be restarted from a stalled fork without formation of a D-loop? PriA preferentially binds to forks with the 3Ј end of a leading strand present at the branch point (32). PriA can also assemble a competent replication complex that can utilize this 3Ј end for priming of replication (11).…”

Section: Discussionmentioning

confidence: 99%

Rescue of stalled replication forks by RecG: Simultaneous translocation on the leading and lagging strand templates supports an active DNA unwinding model of fork reversal and Holliday junction formation

McGlynn

Lloyd

2001

Proc. Natl. Acad. Sci. U.S.A.

173

190

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Modification of damaged replication forks is emerging as a crucial factor for efficient chromosomal duplication and the avoidance of genetic instability. The RecG helicase of Escherichia coli, which is involved in recombination and DNA repair, has been postulated to act on stalled replication forks to promote replication restart via the formation of a four-stranded (Holliday) junction. Here we show that RecG can actively unwind the leading and lagging strand arms of model replication fork structures in vitro. Unwinding is achieved in each case by simultaneous interaction with and translocation along both the leading and lagging strand templates at a fork. Disruption of either of these interactions dramatically inhibits unwinding of the opposing duplex arm. Thus, RecG translocates simultaneously along two DNA strands, one with 5 -3 and the other with 3 -5 polarity. The unwinding of both nascent strands at a damaged fork, and their subsequent annealing to form a Holliday junction, may explain the ability of RecG to promote replication restart. Moreover, the preferential binding of partial forks lacking a leading strand suggests that RecG may have the ability to target stalled replication intermediates in vivo in which lagging strand synthesis has continued beyond the leading strand.DNA repair ͉ recombination ͉ helicases

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“…Here, the DSB generated is processed by RecBCD to generate a recombinogenic 3Ј single-stranded tail that is used for RecAcatalyzed strand invasion with the intact sister duplex, creating a D loop. This structure is recognized by PriA (4,5), which then directs the assembly of a new replication fork at the site through the loading of a primosome (6). The strand crossover initiating D loop formation is resolved subsequently.…”

mentioning

confidence: 99%

Formation of Holliday junctions by regression of nascent DNA in intermediates containing stalled replication forks: RecG stimulates regression even when the DNA is negatively supercoiled

McGlynn

Lloyd

Marians

2001

Proc. Natl. Acad. Sci. U.S.A.

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147

116

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Replication forks formed at bacterial origins often encounter template roadblocks in the form of DNA adducts and frozen protein-DNA complexes, leading to replication-fork stalling and inactivation. Subsequent correction of the corrupting template lesion and origin-independent assembly of a new replisome therefore are required for survival of the bacterium. A number of models for replication-fork restart under these conditions posit that nascent strand regression at the stalled fork generates a Holliday junction that is a substrate for subsequent processing by recombination and repair enzymes. We show here that early replication intermediates containing replication forks stalled in vitro by the accumulation of excess positive supercoils could be cleaved by the Holliday junction resolvases RusA and RuvC. Cleavage by RusA was inhibited by the presence of RuvA and was stimulated by RecG, confirming the presence of Holliday junctions in the replication intermediate and supporting the previous proposal that RecG could catalyze nascent strand regression at stalled replication forks. Furthermore, RecG promoted Holliday junction formation when replication intermediates in which the replisome had been inactivated were negatively supercoiled, suggesting that under intracellular conditions, the action of RecG, or helicases with similar activities, is necessary for the catalysis of nascent strand regression. T he picture of how DNA replication proceeds around the bacterial chromosome has changed over the last decade as a result of research in many laboratories (1). Even though the two replication forks that form at oriC have a sufficiently high enough inherent processivity to complete replication of the chromosome, it is clear that this is generally not what happens. Instead, the replication forks formed at the origin become inactivated at high frequency as a result of an encounter with roadblocks either in or on the template strands. These roadblocks can take many forms: a nick in one of the template strands, a DNA adduct formed as a result of endogenous damage, secondary structure in the template, and frozen proteins on the DNA. Survival then depends on both correction of the damage and reactivation of DNA replication.Although there is a large body of both genetic and biochemical data informing the mechanisms that act to repair damaged nucleotides in DNA, except in one instance, the mechanisms of replication-fork restart are less well defined. Replication-fork restart after an encounter with a template nick, leading to double-strand break (DSB) generation and detachment from the growing fork of one of the nascent sister duplexes-sometimes termed replication fork collapse (2)-is effected by a marriage of homologous recombination and DNA replication proteins (3). Here, the DSB generated is processed by RecBCD to generate a recombinogenic 3Ј single-stranded tail that is used for RecAcatalyzed strand invasion with the intact sister duplex, creating a D loop. This structure is recognized by PriA (4, 5), which then directs the a...

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Two Modes of PriA Binding to DNA

Cited by 91 publications

References 38 publications

The PriA Replication Restart Protein Blocks Replicase Access Prior to Helicase Assembly and Directs Template Specificity through Its ATPase Activity

The PriA Replication Restart Protein Blocks Replicase Access Prior to Helicase Assembly and Directs Template Specificity through Its ATPase Activity

Rescue of stalled replication forks by RecG: Simultaneous translocation on the leading and lagging strand templates supports an active DNA unwinding model of fork reversal and Holliday junction formation

Formation of Holliday junctions by regression of nascent DNA in intermediates containing stalled replication forks: RecG stimulates regression even when the DNA is negatively supercoiled

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