Control of Smc Coiled Coil Architecture by the ATPase Heads Facilitates Targeting to Chromosomal ParB/parS and Release onto Flanking DNA

Minnen, Anita; Bürmann, Frank; Wilhelm, Larissa; Anchimiuk, Anna; Diebold-Durand, Marie-Laure; Gruber, Stephan

doi:10.1016/j.celrep.2016.01.066

Cited by 84 publications

(192 citation statements)

References 47 publications

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“…Interestingly, this localization phenotype is similar to the EQ mutant, which in contrast to the Mini-Smc proteins is blocked in ATP hydrolysis (Hirano and Hirano, 2004, Minnen et al., 2016) (Figures 4A and S4A). We confirmed these findings by ChIP coupled to quantitative PCR (qPCR), which also showed that the extent of redistribution correlated well with the ability of Mini-Smc proteins to support fast growth (Figure S4B).…”

Section: Resultsmentioning

confidence: 71%

“…We performed site-specific cysteine crosslinking experiments in vivo and found that dimerization at the hinge domain was normal in the six tested Mini-Smc proteins (Figures 3A and S3B). Likewise, the Smc head domains engaged robustly in the presence of a mutation blocking ATP hydrolysis (E1118Q [EQ]), whereas head engagement was hardly detectable in the absence of this mutation, as observed with full-length Smc (Minnen et al., 2016) (Figures 3B and S3C; see Figure S3A for an overview of the SMC ATP hydrolysis cycle). Curiously, the efficiency of head engagement in Smc(EQ) proteins increased somewhat as the coiled coil was shortened (Figures 3B and S3D), consistent with our previous proposition that the formation of a rod by close juxtaposition of the two Smc coiled coils hinders head engagement (Minnen et al., 2016, Soh et al., 2015).…”

Section: Resultsmentioning

confidence: 99%

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Tuned SMC Arms Drive Chromosomal Loading of Prokaryotic Condensin

et al. 2017

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SummarySMC proteins support vital cellular processes in all domains of life by organizing chromosomal DNA. They are composed of ATPase “head” and “hinge“ dimerization domains and a connecting coiled-coil “arm.” Binding to a kleisin subunit creates a closed tripartite ring, whose ∼47-nm-long SMC arms act as barrier for DNA entrapment. Here, we uncover another, more active function of the bacterial Smc arm. Using high-throughput genetic engineering, we resized the arm in the range of 6–60 nm and found that it was functional only in specific length regimes following a periodic pattern. Natural SMC sequences reflect these length constraints. Mutants with improper arm length or peptide insertions in the arm efficiently target chromosomal loading sites and hydrolyze ATP but fail to use ATP hydrolysis for relocation onto flanking DNA. We propose that SMC arms implement force transmission upon nucleotide hydrolysis to mediate DNA capture or loop extrusion.

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Section: Resultsmentioning

confidence: 71%

Section: Resultsmentioning

confidence: 99%

Tuned SMC Arms Drive Chromosomal Loading of Prokaryotic Condensin

et al. 2017

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“…ScpAB have been shown to convert Smc—that binds DNA via coiled coil opening at hinge domains (loosely-DNA bound Smc)—to stably DNA-bound Smc rings, which involves head interactions of Smc (62). We show that ∼60% of ScpA monomers (about 50 monomers per cell) are statically positioned, dependent on the presence of Smc, and therefore must be bound to Smc dimers at the SMC clusters, while 40% of ScpA are diffusing through the cell.…”

Section: Discussionmentioning

confidence: 99%

Single molecule tracking reveals that the bacterial SMC complex moves slowly relative to the diffusion of the chromosome

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Borgmann

Rösch

et al. 2018

Nucleic Acids Research

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Structural Maintenance of Chromosomes (SMC) proteins and their complex partners (ScpA and ScpB in many bacteria) are involved in chromosome compaction and segregation in all kinds of organisms. We employed single molecule tracking (SMT), tracking of chromosomal loci, and single molecule counting in Bacillus subtilis to show that in slow growing cells, ∼30 Smc dimers move throughout the chromosome in a constrained mode, while ∼60 ScpA and ScpB molecules travel together in a complex, but independently of the nucleoid. Even an Smc truncation that lacks the ATP binding head domains still scans the chromosome, highlighting the importance of coiled coil arm domains. When forming a complex, 10–15 Smc/ScpAB complexes become essentially immobile, moving slower than chromosomal loci. Contrarily, SMC-like protein RecN, which forms assemblies at DNA double strand breaks, moves faster than chromosome sites. In the absence of Smc, chromosome sites investigated were less mobile than in wild type cells, indicating that Smc contributes to chromosome dynamics. Thus, our data show that Smc/ScpAB clusters occur at several sites on the chromosome and contribute to chromosome movement.

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“…subtilis and Streptococcus pneumoniae , where the ParB- parS complex recruits SMC to the oriC region, thereby allowing correct chromosome segregation [14–16]. This recruitment is thought to depend on ParB ability to bridge DNA, a phenomenon previously described as “spreading” [10], but may also require a specific interaction of SMC with ParB [17,18]. …”

Section: Introductionmentioning

confidence: 99%

Regional Control of Chromosome Segregation in Pseudomonas aeruginosa

2016

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Chromosome segregation in bacteria occurs concomitantly with DNA replication, and the duplicated regions containing the replication origin oriC are generally the first to separate and migrate to their final specific location inside the cell. In numerous bacterial species, a three-component partition machinery called the ParABS system is crucial for chromosome segregation. This is the case in the gammaproteobacterium Pseudomonas aeruginosa, where impairing the ParABS system is very detrimental for growth, as it increases the generation time and leads to the formation of anucleate cells and to oriC mispositioning inside the cell. In this study, we investigate in vivo the ParABS system in P. aeruginosa. Using chromatin immuno-precipitation coupled with high throughput sequencing, we show that ParB binds to four parS site located within 15 kb of oriC in vivo, and that this binding promotes the formation of a high order nucleoprotein complex. We show that one parS site is enough to prevent anucleate cell formation, therefore for correct chromosome segregation. By displacing the parS site from its native position on the chromosome, we demonstrate that parS is the first chromosomal locus to be separated upon DNA replication, which indicates that it is the site of force exertion of the segregation process. We identify a region of approximatively 650 kb surrounding oriC in which the parS site must be positioned for chromosome segregation to proceed correctly, and we called it “competence zone” of the parS site. Mutant strains that have undergone specific genetic rearrangements allow us to propose that the distance between oriC and parS defines this “competence zone”. Implications for the control of chromosome segregation in P. aeruginosa are discussed.

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Control of Smc Coiled Coil Architecture by the ATPase Heads Facilitates Targeting to Chromosomal ParB/parS and Release onto Flanking DNA

Cited by 84 publications

References 47 publications

Tuned SMC Arms Drive Chromosomal Loading of Prokaryotic Condensin

Tuned SMC Arms Drive Chromosomal Loading of Prokaryotic Condensin

Single molecule tracking reveals that the bacterial SMC complex moves slowly relative to the diffusion of the chromosome

Regional Control of Chromosome Segregation in Pseudomonas aeruginosa

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