|
| Status |
Public on May 31, 2023 |
| Title |
Hyperphosphorylation of the Group A Streptococcal Control of Virulence Regulator Increases Promoter Occupancy Specifically at Virulence Factor Encoding Genes |
| Organism |
Streptococcus pyogenes |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
The control of virulence two-component gene regulatory system (CovRS) is critical to the pathogenesis of many medically important streptococci. In emm1 group A streptococci (GAS), CovR directly binds the promoters of numerous GAS virulence factor encoding genes. Elimination of CovS phosphatase activity increases CovR phosphorylation (CovR~P) levels and abrogates GAS virulence. Given the emm type-specific diversity of CovRS function, herein we used ChIP-seq to define global CovR DNA occupancy in the wild-type emm3 strain MGAS10870 (medium CovR~P) and its CovS phosphatase-negative derivative 10870-CovS-T284A (high CovR~P). In the wild-type emm3 strain, 89% of the previously identified emm1 CovR binding sites present in the emm3 genome were also enriched; additionally, we ascertained unique CovR binding, primarily to genes in mobile genetic elements and other sites of inter-strain chromosomal differences. Elimination of phosphatase activity specifically increased CovR occupancy at the promoters of a broad array of CovR repressed virulence factor encoding genes, including those encoding the key GAS regulator Mga and M protein. However, a limited number of promoters had augmented enrichment at low CovR~P levels. Differential motif searches using sequences enriched at high vs. low CovR~P levels revealed two distinct binding patterns. At high CovR~P, a pseudo-palindromic AT-rich consensus sequence consistent with CovR binding as a dimer was determined. Conversely, sequences specifically enriched at low CovR~P contained isolated “ATTARA” motifs suggesting an interaction with a monomer. These data extend understanding of global CovR DNA occupancy beyond emm1 GAS and provide a mechanism for previous observations regarding hypovirulence induced by CovS phosphatase abrogation.
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| Overall design |
ChIP-seq analysis using anti-CovR antibody of wild-type (MGAS10870) and high phosphorylation (10870-CovS-T284A) strains
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| |
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| Contributor(s) |
Horstmann N, Tran CN, Flores AR, Shelburne SA |
| Citation(s) |
37289078 |
| |
| Submission date |
Apr 28, 2023 |
| Last update date |
Aug 30, 2023 |
| Contact name |
William Charles Shropshire |
| E-mail(s) |
wcshropshire@mdanderson.org
|
| Phone |
8307082542
|
| Organization name |
The University of Texas MD Anderson Cancer Center
|
| Street address |
1901 East Road
|
| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77054 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL28678 |
Illumina NovaSeq 6000 (Streptococcus pyogenes) |
|
| Samples (11)
|
| GSM7245938 |
MGAS10870 - input - 5 - input DNA |
| GSM7245939 |
MGAS10870-CovS-T284A - output - 1 - biol rep 1 |
| GSM7245940 |
MGAS10870-CovS-T284A - output - 2 - biol rep 2 |
| GSM7245941 |
MGAS10870-CovS-T284A - output - 5 - biol rep 3 |
| GSM7245942 |
MGAS10870-CovS-T284A - input - 15 - input DNA |
| GSM7245943 |
10870-CovR - output - 3 - biol rep 1 |
| GSM7245944 |
10870-CovR - output - 4 - biol rep 2 |
| GSM7245945 |
10870-CovR - input - 10 - input DNA |
|
| Relations |
| BioProject |
PRJNA962888 |
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