Identification and Analysis of the regulatory RNA TrmS in the probiotic bacteria Streptococcus thermophilus

Presentation Type

Poster

Full Name of Faculty Mentor

Brian M. Lee, Chemistry

Major

Biochemistry

Presentation Abstract

The human microbiome contains billions of bacteria, primarily found in the gut. Many of these bacteria are non-pathogenic and could have a beneficial relationship with our cells. One non-pathogenic species of interest is Streptococcus thermophilus for its frequent use in the dairy industry. To further understand these bacteria, we must study how they regulate essential functions related to survival, reproduction, and potential host interaction. Genes can be regulated at the transcriptional level by regulatory RNA which fold into tertiary structures that can interact with other RNA, proteins, or both. Our goal is to identify and characterize regulatory RNA in S. thermophilus along with their associated proteins to characterize potential communication between probiotic bacteria and host cells. Candidate RNA TrmS was identified based on a potential cis-regulatory element downstream of a tRNALeu methyltransferase in S. thermophilus. Secondary and tertiary structures were predicted. TrmS DNA was successfully isolated and amplified through PCR, and RNA was synthesized through T7 transcription reactions, which was confirmed using gel electrophoresis. Thermal melt assays were performed to confirm the presence of secondary structures, which gave results expected by structure predictions. This allowed for further prediction of the mechanism of interaction between TrmS and the neighboring predicted tRNALeu methyltransferase protein-encoding gene. Alignment of the protein found in S. thermophilus with those of known structure suggests a similar function in methylating the anticodon loop of tRNALeu. TrmS sequence alignments identified conserved loops regions in TrmS across several bacterial species, suggesting potential targets for protein or RNA interaction.

Start Date

12-4-2023 4:00 PM

End Date

12-4-2023 6:00 PM

Disciplines

Biochemistry

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Apr 12th, 4:00 PM Apr 12th, 6:00 PM

Identification and Analysis of the regulatory RNA TrmS in the probiotic bacteria Streptococcus thermophilus

The human microbiome contains billions of bacteria, primarily found in the gut. Many of these bacteria are non-pathogenic and could have a beneficial relationship with our cells. One non-pathogenic species of interest is Streptococcus thermophilus for its frequent use in the dairy industry. To further understand these bacteria, we must study how they regulate essential functions related to survival, reproduction, and potential host interaction. Genes can be regulated at the transcriptional level by regulatory RNA which fold into tertiary structures that can interact with other RNA, proteins, or both. Our goal is to identify and characterize regulatory RNA in S. thermophilus along with their associated proteins to characterize potential communication between probiotic bacteria and host cells. Candidate RNA TrmS was identified based on a potential cis-regulatory element downstream of a tRNALeu methyltransferase in S. thermophilus. Secondary and tertiary structures were predicted. TrmS DNA was successfully isolated and amplified through PCR, and RNA was synthesized through T7 transcription reactions, which was confirmed using gel electrophoresis. Thermal melt assays were performed to confirm the presence of secondary structures, which gave results expected by structure predictions. This allowed for further prediction of the mechanism of interaction between TrmS and the neighboring predicted tRNALeu methyltransferase protein-encoding gene. Alignment of the protein found in S. thermophilus with those of known structure suggests a similar function in methylating the anticodon loop of tRNALeu. TrmS sequence alignments identified conserved loops regions in TrmS across several bacterial species, suggesting potential targets for protein or RNA interaction.