Poster abstracts
Poster number 157 submitted by Chunxi Zeng
Molecular modulation of T box riboswitch function
Chunxi Zeng (Ohio University), Shu Zhou, Jennifer V. Hines (Ohio University)
Abstract:
The T box riboswitch is a transcription regulation mechanism that controls expression of essential genes in many bacteria. Uncharged cognate tRNA induces read-through (antitermination) of the T box riboswitch and allows expression of downstream genes. Two fluorescently labeled probes were designed to study the riboswitch in real time during multi-round in vitro transcription of the leader region of the glyQS gene. The read-through probe showed good photostability, integrity and target sensitivity. Specific riboswitch response to uncharged cognate tRNAgly was detected with a low nanomolar EC50. Using this validated, moderate throughput riboswitch functional assay, the effect of molecular modulators of T box riboswitch function was then determined.
References:
Green, N.J., et al. (2010) The T box mechanism: tRNA as a regulatory molecule. FEBS Lett., 584, 318–24.
Grundy, F.J., et al. (2002) tRNA-mediated transcription antitermination in vitro: codon-anticodon pairing independent of the ribosome. PNAS, 99, 11121–6.
Putzer, H., et al. (2002) Transfer RNA-mediated antitermination in vitro. Nucleic Acids Res., 30, 3026–33.
Tsourkas, A., et al. (2002) Hybridization of 2’-O-methyl and 2'-deoxy molecular beacons to RNA and DNA targets. Nucleic Acids Res., 30, 5168–5174.
Nakano, S., et al. (2009) Facilitation of RNA enzyme activity in the molecular crowding media of cosolutes. J. Am. Chem. Soc., 131, 16881–8.
Keywords: T box riboswitch, molecular beacon, in vitro transcription