Using a riboswitch sensor to examine coenzyme B(12) metabolism and transport in E. coli.
Small molecules play crucial roles in every major cellular process. Despite this, detecting their levels within cells remains a significant challenge. Here, we describe intracellular sensors of coenzyme B(12) that make use of the exquisite molecular detection capabilities of a naturally occurring riboswitch. These probes sensitively detect their target using colorimetric, fluorescent, or luminescent reporters. To assess their utility in the study of biological systems, the sensors were applied to examine the synthesis and the import of coenzyme B(12). The sensors were able to monitor the effects of genetic deletions, recombinant expression of foreign genes, and varied growth conditions on both of these processes. These results indicate that riboswitch-based sensors can provide valuable information on intracellular small molecule concentrations that can be employed in the study of related cellular processes.
Fowler
Casey C
CC
Department of Biochemistry and Biomedical Sciences, McMaster University, 1200 Main Street West, Hamilton, ON L8N3Z5, Canada.
Brown
Eric D
ED
Li
Yingfu
Y
D001692:Biological Transport; D015374:Biosensing Techniques / Q000379:methods*; D003038:Cobamides / Q000378:metabolism*; D004926:Escherichia coli / Q000166:cytology / Q000378:metabolism*; D008024:Ligands; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D038621:Regulatory Sequences, Ribonucleic Acid; D013997:Time Factors
D003038:Cobamides; D008024:Ligands; D012333:RNA, Messenger; D038621:Regulatory Sequences, Ribonucleic Acid