Fast, DNA-sequence independent translocation by FtsK in a single-molecule experiment

Saleh, Omar A.; Perals, Corine; Barre, François‐Xavier; Allemand, Jean-François

doi:10.1038/sj.emboj.7600242

Cited by 134 publications

(172 citation statements)

References 39 publications

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“…The N-terminus is responsible for targeting FtsK to the division machinery, where it plays an essential role in the septation process Yu et al, 1998]. The Cterminus, by contrast, is dispensable for divisiome function and acts as an ATP-dependent DNA translocase [Aussel et al, 2002;Saleh et al, 2004] that pumps DNA away from the septal region of the cells [Liu et al, 1998;Yu et al, 1998]. It is still unclear how FtsK determines the direction its substrate has to be translocated to be positioned in the correct daughter cell compartment.…”

Section: The Last Stages Of Dna Replicationmentioning

confidence: 99%

The bacterial nucleoid: A highly organized and dynamic structure

Thanbichler

Wang

Shapiro

2005

J of Cellular Biochemistry

122

101

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Recent advances in bacterial cell biology have revealed unanticipated structural and functional complexity, reminiscent of eukaryotic cells. Particular progress has been made in understanding the structure, replication, and segregation of the bacterial chromosome. It emerged that multiple mechanisms cooperate to establish a dynamic assembly of supercoiled domains, which are stacked in consecutive order to adopt a defined higher-level organization. The position of genetic loci on the chromosome is thereby linearly correlated with their position in the cell. SMC complexes and histone-like proteins continuously remodel the nucleoid to reconcile chromatin compaction with DNA replication and gene regulation. Moreover, active transport processes ensure the efficient segregation of sister chromosomes and the faithful restoration of nucleoid organization while DNA replication and condensation are in progress.

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Section: The Last Stages Of Dna Replicationmentioning

confidence: 99%

The bacterial nucleoid: A highly organized and dynamic structure

Thanbichler

Wang

Shapiro

2005

J of Cellular Biochemistry

122

101

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“…SpoIIIE homologues are widespread in non-sporulating bacteria, including the conserved cell division protein FtsK. FtsK probably functions like SpoIIIE to ensure the fidelity of chromosome segregation into the daughter cells prior to septation (Aussel et al 2002), and has been shown in a recent single molecule experiment to act as a DNA translocase in vitro (Saleh et al 2004).…”

Section: Orientation Positioning and Movement Of The Bacterial Chrommentioning

confidence: 99%

Towards understanding the molecular basis of bacterial DNA segregation

Leonard

Møller‐Jensen

Löwe

2005

Phil. Trans. R. Soc. B

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Bacteria ensure the fidelity of genetic inheritance by the coordinated control of chromosome segregation and cell division. Here, we review the molecules and mechanisms that govern the correct subcellular positioning and rapid separation of newly replicated chromosomes and plasmids towards the cell poles and, significantly, the emergence of mitotic-like machineries capable of segregating plasmid DNA. We further describe surprising similarities between proteins involved in DNA partitioning (ParA/ParB) and control of cell division (MinD/MinE), suggesting a mechanism for intracellular positioning common to the two processes. Finally, we discuss the role that the bacterial cytoskeleton plays in DNA partitioning and the missing link between prokaryotes and eukaryotes that is bacterial mechano-chemical motor proteins.

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“…However, single-molecule observations of the FtsK motor domain (FtsK 50C ) translocating on DNA showed that FtsK could spontaneously reverse direction while maintaining an overall directionality (5,10). Our prior study implies that recurring DNA sequences regulate the direction of FtsK translocation (5).…”

mentioning

confidence: 99%

Identification of oligonucleotide sequences that direct the movement of the Escherichia coli FtsK translocase

Levy

Ptacin²,

Pease³

et al. 2005

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

110

116

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FtsK from Escherichia coli is a fast and sequence-directed DNA translocase with roles in chromosome dimer resolution, segregation, and decatenation. From the movement of single FtsK particles on defined DNA substrates and an analysis of skewed DNA sequences in bacteria, we identify GNGNAGGG, its complement, or both as a sequence motif that controls translocation directionality. GNGNAGGG is skewed so that it is predominantly on the leading strand of chromosomal replication. Translocation across this octamer from the 3 side of the G-rich strand causes FtsK to pause, turn around, and translocate in the opposite direction. Only 39 ؎ 4% of the encounters between FtsK and the octamer result in a turnaround, congruent with our optimum turnaround probability prediction of 30%. The probability that the observed skew of GNGNAGGG within 1 megabase of dif occurred by chance in E. coli is 1.7 ؋ 10 ؊57 , and similarly dramatic skews are found in the five other bacterial genomes we examined. The fact that FtsK acts only in the terminus region and the octamer skew extends from origin to terminus implies that this skew is also important in other basic cellular processes that are common among bacteria. Finally, we show that the FtsK translocase is a powerful motor that is able to displace a triplex-forming oligo from a DNA substrate.dimer resolution ͉ sequence-directed translocases ͉ skewed sequences ͉ triplex displacement ͉ single-molecule T he DNA sequence of an organism encodes information on multiple levels. Beyond directly encoding proteins, specific DNA sequences recruit and direct the proteins required for processes such as DNA replication, transcription, and repair. DNA sequence-dependent proteins typically locate their binding site by diffusion where they then carry out a specific action (1). In contrast, the Escherichia coli DNA translocase FtsK uses an ATP-powered search mechanism to reach its target (dif ), the site of chromosome dimer resolution (2-5). During replication, crossing over by homologous recombination can lead to the formation of a chromosome dimer that must be resolved into monomers before segregation (2). FtsK is required to activate the site-specific recombinases XerC and XerD that resolve these dimers at the dif site, which is near the terminus of replication (2, 6). FtsK is anchored in the membrane at the division septum and actively translocates the dif sites into spatial proximity, rather than waiting for dif to diffuse into the septal region (7-9). The overall efficiency of reaching dif by translocation is greatly affected by two factors. First, FtsK must be able to bypass or displace bound proteins or other potential roadblocks it may encounter on the DNA. Second, FtsK must maintain its overall direction of translocation toward dif.Perhaps the simplest mechanism for ensuring that FtsK always translocates toward dif would involve FtsK loading at sequences that point FtsK in the proper direction. However, single-molecule observations of the FtsK motor domain (FtsK 50C ) translocating on DNA showed ...

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Fast, DNA-sequence independent translocation by FtsK in a single-molecule experiment

Cited by 134 publications

References 39 publications

The bacterial nucleoid: A highly organized and dynamic structure

The bacterial nucleoid: A highly organized and dynamic structure

Towards understanding the molecular basis of bacterial DNA segregation

Identification of oligonucleotide sequences that direct the movement of the Escherichia coli FtsK translocase

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