Identification of a putative RocS homolog through phenotypic profiling of uncharacterized essential genes in Streptococcus mutans
- 1University of Florida College of Dentistry
Abstract
Genome-wide viability catalogs produced by transposon sequencing (Tn-seq) and CRISPR interference (CRISPRi) have successfully mapped the essential genome of Streptococcus mutans. In this study, we combined predictive bioinformatics, conditional CRISPRi transcriptional silencing, transmission electron microscopy, transcriptomics, and genetic suppressor screens to investigate nine poorly characterized essential genes in S. mutans. From this screen, phenotypic and genetic analyses identified SMU_393 as a functional homolog of the pneumococcal chromosome segregation factor, RocS. Depletion of SMU_393 resulted in abnormal cell widening, hypersensitivity to DNA damage, and a significant subpopulation of anucleate cells. These phenotypes were bypassed by a spontaneous surface-exposed missense mutation (dnaAQ197E) within the AAA+ ATPase domain of the replication initiator. Together, this study refines annotations within the S. mutans essential genome and provides genetic insights into streptococcal chromosome segregation and cell cycle control.
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BibTeX
@misc{dover2026identification,
title = {{Identification of a putative RocS homolog through phenotypic profiling of uncharacterized essential genes in Streptococcus mutans}},
author = {C. Dover and K. Tamrakar and B. Dwivedi and E. R. Roberts and S. Chudal and S. King and E. S. Chavez and V. de Crecy-Lagard and R. C. Shields},
howpublished = {bioRxiv},
year = {2026},
month = jul,
doi = {10.64898/2026.07.15.738700},
url = {https://www.biorxiv.org/content/10.64898/2026.07.15.738700v2},
note = {CC BY},
}
RIS
TY - UNPB
TI - Identification of a putative RocS homolog through phenotypic profiling of uncharacterized essential genes in Streptococcus mutans
AU - C. Dover
AU - K. Tamrakar
AU - B. Dwivedi
AU - E. R. Roberts
AU - S. Chudal
AU - S. King
AU - E. S. Chavez
AU - V. de Crecy-Lagard
AU - R. C. Shields
PY - 2026
DA - 2026/07/15
PB - bioRxiv
DO - 10.64898/2026.07.15.738700
UR - https://www.biorxiv.org/content/10.64898/2026.07.15.738700v2
AB - Genome-wide viability catalogs produced by transposon sequencing (Tn-seq) and CRISPR interference (CRISPRi) have successfully mapped the essential genome of Streptococcus mutans. In this study, we combined predictive bioinformatics, conditional CRISPRi transcriptional silencing, transmission electron microscopy, transcriptomics, and genetic suppressor screens to investigate nine poorly characterized essential genes in S. mutans. From this screen, phenotypic and genetic analyses identified SMU_393 as a functional homolog of the pneumococcal chromosome segregation factor, RocS. Depletion of SMU_393 resulted in abnormal cell widening, hypersensitivity to DNA damage, and a significant subpopulation of anucleate cells. These phenotypes were bypassed by a spontaneous surface-exposed missense mutation (dnaAQ197E) within the AAA+ ATPase domain of the replication initiator. Together, this study refines annotations within the S. mutans essential genome and provides genetic insights into streptococcal chromosome segregation and cell cycle control.
ER -