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Abstract Research on chromosome organization and cell cycle progression in spherical bacteria, particularlyStaphylococcus aureus, remains limited and fragmented. In this study, we established a working model to investigate chromosome dynamics inS. aureususing a Fluorescent Repressor-Operator System (FROS), which enabled precise localization of specific chromosomal loci. This approach revealed that theS. aureuscell cycle and chromosome replication cycle are not coupled, with cells exhibiting two segregated origins of replication at the start of the cell cycle. The chromosome has a specific origin-terminus-origin conformation, with origins localizing near the membrane, towards the tip of each hemisphere, or the “cell poles”. We further used this system to assess the role of various proteins with a role inS. aureuschromosome biology, focusing on the ParB-parSand SMC-ScpAB systems. Our results demonstrate that ParB binds fiveparSchromosomal sequences and the resulting complexes influence chromosome conformation, but play a minor role in chromosome compaction and segregation. In contrast, the SMC-ScpAB complex plays a key role inS. aureuschromosome biology, contributing to chromosome compaction, segregation and spatial organization. Additionally, we systematically assessed and compared the impact of proteins linking chromosome segregation to cell division—Noc, FtsK, SpoIIIE and XerC—on origin and terminus number and positioning. This work provides a comprehensive study of the factors governing chromosome dynamics and organization inS. aureus, contributing to our knowledge on chromosome biology of spherical bacteria.
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This content will become publicly available on February 23, 2026
Cell size regulation in bacteria: a tale of old regulators with new mechanisms
SUMMARY Proper function in a bacterial cell relies on intrinsic cell size regulation. The molecular mechanisms underlying how bacteria maintain their cell size remain unclear. The conserved regulator DnaA, the initiator of chromosome replication, is associated to size regulation by controlling the number of origins of replication (oriC) per cell. In this study, we identify and characterize a new mechanism in which DnaA modulates cell size independently oforiC-copy number. By altering the levels of DnaA without impacting chromosome replication, we demonstrate that DnaA’s activity as a transcription factor can slow down cell elongation rate resulting in cells that are ∼20% smaller. We identify the peptidoglycan biosynthetic enzyme MurD as a key player of cell size regulation inCaulobacter crescentusand in the evolutionarily distant bacteriumEscherichia coli. Collectively, our findings provide mechanistic insights to the complex regulation of cell size in bacteria.
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- Award ID(s):
- 2243257
- PAR ID:
- 10595347
- Publisher / Repository:
- bioRxiv
- Date Published:
- Format(s):
- Medium: X
- Institution:
- bioRxiv
- Sponsoring Org:
- National Science Foundation
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