In bacteria, chromosomal DNA resides in the cytoplasm, and most transcription factors are also found in the cytoplasm. However, some transcription factors, called membrane-bound transcription factors (MTFs), reside in the cytoplasmic membrane. Here, we report the identification of a new MTF in the Gram-positive pathogen Staphylococcus aureus and its regulation by the protease FtsH. The MTF, named MbtS (membrane-bound transcription factor of Staphylococcus aureus), is encoded by SAUSA300_2640 and predicted to have an N-terminal DNA binding domain and three transmembrane helices. The MbtS protein was degraded by membrane vesicles containing FtsH or by the purified FtsH. MbtS bound to an inverted repeat sequence in its promoter region, and the DNA binding was essential for its transcription. Transcriptional comparison between the ftsH deletion mutant and the ftsH mbtS double mutant showed that MbtS could alter the transcription of over 200 genes. Although the MbtS protein was not detected in wild-type (WT) cells grown in a liquid medium, the protein was detected in some isolated colonies on an agar plate. In a murine model of a skin infection, the disruption of mbtS increased the lesion size. Based on these results, we concluded that MbtS is a new S. aureus MTF whose activity is proteolytically regulated by FtsH. IMPORTANCE Staphylococcus aureus is an important pathogenic bacterium causing various diseases in humans. In the bacterium, transcription is typically regulated by the transcription factors located in the cytoplasm. In this study, we report an atypical transcription factor identified in S. aureus. Unlike most other transcription factors, the newly identified transcription factor is located in the cytoplasmic membrane, and its activity is proteolytically controlled by the membrane-bound AAA+ protease FtsH. The newly identified MTF, named MbtS, has the potential to regulate the transcription of over 200 genes. This study provides a molecular mechanism by which a protease affects bacterial transcription and illustrates the diversity of the bacterial transcriptional regulation.
Background: Fights, bullying, and gun violence in the Northwest Indiana and Chicagoland middle schools and high schools can have long lasting effects on a student’s physical, emotional, or mental wellbeing. Project Outreach and Prevention on Youth Violence (henceforth POP) is a foundation started by two NWI doctors, Dr. Michael McGee and Dr. Reuban Rutland, looking to do their part in preventing victims of youth violence from reaching their emergency departments. POP also reaches students underrepresented in medicine to breach that gap. Programs such as POP’s Health Professions Enrichment Program (HPEP). POP also utilizes school rallies and violence prevention seminars throughout the year to reach at risk youth. Activities at these rallies include Stop the Bleed training, where students are trained and certified by professionals on how to stop a bleed to various parts of the body. POP also utilizes local celebrities, artists, public figures, and community leaders to incentivize the participation of their target population. Does implementation of anti-violence events with school age students by professionals and community leaders decrease incidences of violence or increase interest in college/STEM professions? Hypothesis: There is an increase in the interest in college/STEM professions. There is a decrease in incidences of violence at schools with active SAVE chapters or participating in an anti-violence event Experimental Design or Project Methods: Surveys, Observations, Student Interviews, Intervention, Cohort Study, Program evaluations, Longitudinal Study (Go to each school nearby and request the info) The study’s population includes students from 7th grade through high school seniors located throughout 13 schools in the northwest Indiana region. POP Results: Analyze Survey Data and interviews for positive correlation with the data. Conclusion/Potential Impact: Pending Review; The potential impact this project yielding positive results would help demonstrate efficacy of POP and enable us to seek more funding opportunities and implement these programs on a wider scale and hopefully being able to see lower instances of violence in areas where programing takes place.
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