Molecular Analysis of pSK1 par: A Novel Plasmid Partitioning System Encoded by Staphylococcal Multiresistance Plasmids

Chan, Helena; Jensen, Slade O.; LeBard, Rebecca; Figgett, William A.; Lai, Evelyn; Simpson, Alice E.; Brzoska, Anthony J.; Davies, Danielle; Connolly, Angela; Cordwell, Stuart J.; Travis, B.A.; Salinas, Raul; Skurray, Ronald A.; Firth, Neville; Schumacher, Maria A.

doi:10.1016/j.jmb.2022.167770

Cited by 7 publications

(5 citation statements)

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“…According to the structure of the corresponding protein-DNA complexes, the helix α2 is inserted in the major groove while the wing primarily contacts the phosphate backbone and possibly the minor groove (30,31). This interaction is also suggested by the Alphafold3 modeling of a RocS dimer in complex with a double-stranded (ds) DNA fragment (Fig.…”

Section: The Dna-binding Domain Of Rocs Is Phosphorylated By Stkpmentioning

confidence: 78%

“…S1) suggests that it could also bind to the membrane and share a common DNA partition mechanism with RocS. While more work is required to identify such mechanism, it can narrow-down previously suggested scenarios (31). By analogy with the partitioning systems of eukaryotic extrachromosomal elements (34), it has been proposed that the Par-dependent plasmid segregation system could bind both specific sequence repeats on the plasmid and degenerated sequence repeats on the chromosome, thus using the nucleoid as a vehicle (31).…”

Section: Discussionmentioning

confidence: 98%

“…3A). Both proteins contain three domains: an N-terminal wHTH domain, a central coiled-coil region, involved in oligomerization and possibly protein-protein interactions, and a C-terminal disordered domain (30,31). RacA contributes to DNA segregation during sporulation through the binding of repetitive centromere-like sequences on the nucleoid and the interaction with the polar anchor DivIVA (4,29,30,33).…”

Section: Discussionmentioning

confidence: 99%

“…While more work is required to identify such mechanism, it can narrow-down previously suggested scenarios (31). By analogy with the partitioning systems of eukaryotic extrachromosomal elements (34), it has been proposed that the Par-dependent plasmid segregation system could bind both specific sequence repeats on the plasmid and degenerated sequence repeats on the chromosome, thus using the nucleoid as a vehicle (31). Therefore, the Par protein could be considered as a sub-cellular positioning system that tethers plasmids to the nucleoid.…”

Section: Discussionmentioning

confidence: 99%

“…Interestingly, a DALI search of the wHTH prediction model of RocS revealed that this domain shares structural homology with the Staphylococcus aureus plasmid partitioning protein Par (28) and the Bacillus subtilis chromosome segregation protein RacA (4, 29) (Fig. 3A), which both harbor a MerR-type wHTH domain (30,31).…”

Section: The Dna-binding Domain Of Rocs Is Phosphorylated By Stkpmentioning

confidence: 99%

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Molecular dissection of the chromosome partitioning protein RocS and regulation by phosphorylation

Demuysere,

Ducret,

Grangeasse

2024

Preprint

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Chromosome segregation in bacteria is a critical process ensuring that each daughter cell receives an accurate copy of the genetic material during cell division. Active segregation factors such as the ParABS system or SMC complexes, are usually essential for this process but are surprisingly dispensable inStreptococcus pneumoniae. Rather, chromosome segregation inS. pneumoniaerelies on the protein RocS, although the molecular mechanisms involved remain elusive. By combining genetics, in vivo imaging and biochemical approaches, we dissected the molecular features of RocS involved in chromosome segregation. Specifically, we investigated the respective function of the three RocS domains, specifically the C-terminal amphipathic helix (AH), the N-terminal DNA-binding domain (DBD) and the coiled-coil domain (CCD) separating the AH and the DBD. Notably, we found that a single AH is not sufficient for membrane binding and that RocS requires prior oligomerization to interact with the membrane. We further demonstrated that this self-interaction was driven by the N-terminal part of the CCD. On the other hand, we revealed that the C-terminal part of the CCD corresponds to a domain of unknown function (DUF 536) defined by three conserved glutamines which play a crucial role in RocS-mediated chromosome segregation. Finally, we showed that the DBD is phosphorylated by the unique serine-threonine kinase ofS. pneumoniaeStkP, and that mimicking this phosphorylation abrogated RocS binding to DNA. Overall, this study offers new insights into chromosome segregation in Streptococci and paves the way for a deeper understanding of RocS-like proteins in other bacteria.

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Section: The Dna-binding Domain Of Rocs Is Phosphorylated By Stkpmentioning

confidence: 78%