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Precursor-product discrimination by La protein during tRNA metabolism

Nature Structural & Molecular Biology volume 16, pages 430–437 (2009)Cite this article

Abstract

La proteins bind pre-tRNAs at their UUU-3′OH ends, facilitating their maturation. Although the mechanism by which La binds pre-tRNA 3′ trailers is known, the function of the RNA binding β-sheet surface of the RNA-recognition motif (RRM1) is unknown. How La dissociates from UUU-3′OH–containing trailers after 3′ processing is also unknown. Here we show that La preferentially binds pre-tRNAs over processed tRNAs or 3′ trailer products through coupled use of two sites: one on the La motif and another on the RRM1 β-surface that binds elsewhere on tRNA. Two sites provide stable pre-tRNA binding, whereas the processed tRNA and 3′ trailer are released from their single sites relatively fast. RRM1 loop-3 mutations decrease affinity for pre-tRNA and tRNA, but not for the UUU-3′OH trailer, and impair tRNA maturation in vivo. We propose that RRM1 functions in activities that are more complex than UUU-3′OH binding. Accordingly, the RRM1 mutations also impair an RNA chaperone activity of La. The results suggest how La distinguishes precursor from product RNAs, allowing it to recycle onto a new pre-tRNA.

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Figure 1: La can bind non–UUU-3′OH–containing RNA via contacts that are not mediated by the previously characterized RNA 3′OH binding site in the La motif.
Figure 2: Two distinct RNA binding sites on La together enhance stable binding to pre-tRNA.
Figure 3: La shows strong preference for pre-tRNA over a UUU-3′OH trailer.
Figure 4: La RRM1 loop-3 mediates UUU-3′OH–independent tRNA binding.
Figure 5: The La loop mutant is defective in tRNA maturation in vivo.
Figure 6: Model of involvement of La protein in a tRNA maturation pathway.

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Acknowledgements

We thank D. Setzer for advice, protocols and RNA binding data analysis tools, M. Nashimoto (Niigata University of Pharmacy and Applied Life Sciences) for the human tRNAArgACG gene and R. Schroeder (University of Vienna) for cis-splicing intron DNA. We thank D. Setzer, D. Engelke and M. Teplova for comments. This work was supported by the Intramural Research Program of the US National Institute of Child Health and Human Development, National Institutes of Health.

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  1. Mark A Bayfield

    Present address: Present address: Department of Biology, York University, Toronto, ON, Canada.,

Authors and Affiliations

  1. Intramural Research Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA

    Mark A Bayfield & Richard J Maraia

  2. Commissioned Corps, US Public Health Service, Bethesda, Maryland, USA

    Richard J Maraia

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M.A.B. performed all experiments; R.J.M. and M.A.B. designed the study and analyzed the data; R.J.M. wrote the paper with editing by M.A.B.

Note: Supplementary information is available on the Nature Structural & Molecular Biology website.

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Correspondence to Richard J Maraia.

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Bayfield, M., Maraia, R. Precursor-product discrimination by La protein during tRNA metabolism. Nat Struct Mol Biol 16, 430–437 (2009). https://doi.org/10.1038/nsmb.1573

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