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RNA Chaperones pp 107-120 | Cite as

Use of tRNA-Mediated Suppression to Assess RNA Chaperone Function

  • Jennifer Porat
  • Mark A. BayfieldEmail author
Protocol
  • 262 Downloads
Part of the Methods in Molecular Biology book series (MIMB, volume 2106)

Abstract

La proteins have well-established roles in the maturation of RNA polymerase III transcripts, including pre-tRNAs. In addition to protecting the 3′ end of pre-tRNAs from exonuclease digestion, La proteins also promote the native fold of the pre-tRNA using RNA chaperone activity. tRNA-mediated suppression in the fission yeast S. pombe has been an invaluable tool in determining the mechanistic basis by which La proteins promote the maturation of defective pre-tRNAs that benefit from RNA chaperone activity. More recently, tRNA-mediated suppression has been adapted to test for RNA chaperone function in the La-related proteins and in the promoting of tRNA function by tRNA modification enzymes. Thus tRNA-mediated suppression can be a useful assay for the investigation of various proteins hypothesized to promote tRNA folding through RNA chaperone related activities.

Key words

RNA chaperone assay Yeast tRNA processing Nonsense suppression Folding Northern blotting Lithium acetate transformation La and La-related proteins (LARPs) tRNA modifications 3′ end protection 

Notes

Acknowledgments

We thank R. Maraia, R. Intine, Y. Huang, A. Sakulich, S. Koduru, S. Hasson, A. Mozlin, J. Mazeika, R. Hussain, A. Vakiloroayaei, and A. Naeeni for their assistance in the development of tRNA-mediated suppression and its use in assessment of RNA chaperone function. We also thank R. Maraia for critical reading of the manuscript. This work is supported by a Discovery Grant from the Natural Science and Engineering Research Council (NSERC) of Canada to MA Bayfield and an NSERC CGS-M scholarship to J Porat.

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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Department of BiologyYork UniversityTorontoCanada

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