Publications by authors named "Malgorzata Michnicka"

2 Publications

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Hierarchical mechanism of amino acid sensing by the T-box riboswitch.

Nat Commun 2018 05 14;9(1):1896. Epub 2018 May 14.

Single Molecule Analysis Group, Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA.

In Gram-positive bacteria, T-box riboswitches control gene expression to maintain the cellular pools of aminoacylated tRNAs essential for protein biosynthesis. Co-transcriptional binding of an uncharged tRNA to the riboswitch stabilizes an antiterminator, allowing transcription read-through, whereas an aminoacylated tRNA does not. Recent structural studies have resolved two contact points between tRNA and Stem-I in the 5' half of the T-box riboswitch, but little is known about the mechanism empowering transcriptional control by a small, distal aminoacyl modification. Using single-molecule fluorescence microscopy, we have probed the kinetic and structural underpinnings of tRNA binding to a glycyl T-box riboswitch. We observe a two-step mechanism where fast, dynamic recruitment of tRNA by Stem-I is followed by ultra-stable anchoring by the downstream antiterminator, but only without aminoacylation. Our results support a hierarchical sensing mechanism wherein dynamic global binding of the tRNA body is followed by localized readout of its aminoacylation status by snap-lock-based trapping.
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http://dx.doi.org/10.1038/s41467-018-04305-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5951919PMC
May 2018

Capture and Release of tRNA by the T-Loop Receptor in the Function of the T-Box Riboswitch.

Biochemistry 2017 07 3;56(28):3549-3558. Epub 2017 Jul 3.

Department of BioSciences, Rice University , Houston, Texas 77005, United States.

In Gram-positive bacteria, the tRNA-dependent T-box riboswitch system regulates expression of amino acid biosynthetic and aminoacyl-tRNA synthetase genes through a transcription attenuation mechanism. Binding of uncharged tRNA "closes" the switch, allowing transcription read-through. Structural studies of the 100-nucleotide stem I domain reveal tRNA utilizes base pairing and stacking interactions to bind the stem, but little is known structurally about the 180-nucleotide riboswitch core (stem I, stem III, and antiterminator stem) in complex with tRNA or the mechanism of coupling of the intermolecular binding domains crucial to T-box function. Here we utilize solution structural and biophysical methods to characterize the interplay of the different riboswitch-tRNA contact points using Bacillus subtilis and Oceanobacillus iheyensis glycyl T-box and T-box:tRNA constructs. The data reveal that tRNA:riboswitch core binding at equilibrium involves only Specifier-anticodon and antiterminator-acceptor stem pairing. The elbow:platform stacking interaction observed in studies of the T-box stem I domain is released after pairing between the acceptor stem and the bulge in the antiterminator helix. The results are consistent with the model of T-box riboswitch:tRNA function in which tRNA is captured by stem I of the nascent mRNA followed by stabilization of the antiterminator helix and the paused transcription complex.
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http://dx.doi.org/10.1021/acs.biochem.7b00284DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5813812PMC
July 2017
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