Presentation Type

Poster

Full Name of Faculty Mentor

Brian M. Lee, Chemistry; Gabriela C. Pérez Alvarado, Chemistry; Sasha S. Bronovitskiy, Georgia Institute of Technology; Finn K. Rose

Major

Biology

Presentation Abstract

The gut microbiome is a complex habitat with many bacterial species. These bacteria play vital roles in regulating several physiological processes in the body. With a rise in the use of probiotics to combat human disease, it is important to understand the mechanisms by which probiotic bacteria regulate host interactions. Our exploration of the physiological functions of probiotic bacteria hopes to elucidate the role of small regulatory RNAs or non-coding RNAs in regulating gene expression within the human body. The goal of this project was to identify and explore the conservation of function and structure of the sRNA, UspS, in two probiotic bacteria and to further analyze its role in host interactions. A candidate sRNA in Lactobacillus species was chosen based on its association with a downstream universal stress protein and conservation among other Lactobacillus species. The sequence of two bacterial species were characterized using computational methods to predict secondary structure, tertiary structure, and mRNA interactions of UspS. Genes for UspS were isolated from two Lactobacillus species, and target sRNAs were synthesized by in vitro transcription using a T7 RNA polymerase. Sequence alignment, secondary, and tertiary structure predictions show conserved pseudoknot region of the P4 region of UspS that may correspond to 6S RNA found in E. coli. 6S RNA is a non-coding RNA that regulates the expression of the sigma subunit of RNA polymerase under stress-related conditions. A thermal melt assay confirmed the presence of secondary structure. Future work will be focused on UspS’ potential role in regulating the expression of the downstream universal stress protein.

Start Date

11-4-2023 10:00 AM

End Date

11-4-2023 12:00 PM

Disciplines

Biology

Included in

Biology Commons

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Apr 11th, 10:00 AM Apr 11th, 12:00 PM

Identification of a Small Regulatory RNA UspS Associated with the Universal Stress Protein in Lactobacillus Species

The gut microbiome is a complex habitat with many bacterial species. These bacteria play vital roles in regulating several physiological processes in the body. With a rise in the use of probiotics to combat human disease, it is important to understand the mechanisms by which probiotic bacteria regulate host interactions. Our exploration of the physiological functions of probiotic bacteria hopes to elucidate the role of small regulatory RNAs or non-coding RNAs in regulating gene expression within the human body. The goal of this project was to identify and explore the conservation of function and structure of the sRNA, UspS, in two probiotic bacteria and to further analyze its role in host interactions. A candidate sRNA in Lactobacillus species was chosen based on its association with a downstream universal stress protein and conservation among other Lactobacillus species. The sequence of two bacterial species were characterized using computational methods to predict secondary structure, tertiary structure, and mRNA interactions of UspS. Genes for UspS were isolated from two Lactobacillus species, and target sRNAs were synthesized by in vitro transcription using a T7 RNA polymerase. Sequence alignment, secondary, and tertiary structure predictions show conserved pseudoknot region of the P4 region of UspS that may correspond to 6S RNA found in E. coli. 6S RNA is a non-coding RNA that regulates the expression of the sigma subunit of RNA polymerase under stress-related conditions. A thermal melt assay confirmed the presence of secondary structure. Future work will be focused on UspS’ potential role in regulating the expression of the downstream universal stress protein.

 

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