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Functional Validation of SAM Riboswitch Element A from Listeria monocytogenes.
Ian Hall1, Kaitlyn Zablock1, Raeleen Sobetski2
1Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States.
The SreA riboswitch in Listeria monocytogenes controls methionine import genes. It functions similarly to other SAM-I riboswitches but at lower concentrations and allows some gene expression even with high SAM levels.
Area of Science:
- Microbiology
- Molecular Biology
- Biochemistry
Background:
- Listeria monocytogenes is an opportunistic foodborne pathogen.
- SreA is a candidate SAM-I riboswitch regulating methionine import genes.
- Riboswitches are crucial for bacterial metabolic regulation.
Purpose of the Study:
- To validate SreA as a functional SAM-I riboswitch.
- To characterize its ligand binding and transcriptional regulation.
- To understand methionine homeostasis in L. monocytogenes.
Main Methods:
- Ligand binding assays
- Structure characterization
- In vitro transcription termination assays
Main Results:
- SreA exhibits structural and binding properties similar to known SAM-I riboswitches.
- SreA induces transcription termination at nanomolar SAM concentrations.
- SreA is a leaky riboswitch, allowing partial gene expression even with millimolar SAM.
Conclusions:
- SreA is a functional SAM-I riboswitch in L. monocytogenes.
- It regulates methionine import genes with high sensitivity to SAM.
- L. monocytogenes likely dampens, rather than shuts off, methionine import gene expression.
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