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Separate RNA-binding surfaces on the multifunctional La protein mediate distinguishable activities in tRNA maturation

Nature Structural & Molecular Biology volume 13, pages 611–618 (2006)Cite this article

Abstract

By sequence-specific binding to 3′ UUU-OH, the La protein shields precursor (pre)-RNAs from 3′ end digestion and is required to protect defective pre–transfer RNAs from decay. Although La is comprised of a La motif and an RNA-recognition motif (RRM), a recent structure indicates that the RRM β-sheet surface is not involved in UUU-OH recognition, raising questions as to its function. Progressively defective suppressor tRNAs in Schizosaccharomyces pombe reveal differential sensitivities to La and Rrp6p, a 3′ exonuclease component of pre-tRNA decay. 3′ end protection is compromised by mutations to the La motif but not the RRM surface. The most defective pre-tRNAs require a second activity of La, in addition to 3′ protection, that requires an intact RRM surface. The two activities of La in tRNA maturation map to its two conserved RNA-binding surfaces and suggest a modular model that has implications for its other ligands.

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Figure 1: Pre–tRNA-M succumbs to rrp6+-mediated nuclear surveillance in S. pombe.
Figure 2: Different sensitivities of several tRNAUGASer alleles to sla1+ and rrp6+.
Figure 3: La mutations examined in this study.
Figure 4: RRM1 β-sheet surface mutations distinguish a second activity of La that is required for defective pre-tRNAUGASer maturation in vivo.
Figure 5: La RRM1 β-sheet surface mutants are active for pre-tRNA 3′ end protection and support La-dependent processing in vivo.
Figure 6: La RRM1 β-sheet surface mutations do not impair high-affinity UUU-OH binding to a short oligo-RNA or pre-tRNA.

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Acknowledgements

We thank M. Blum for media preparation, S. Hasson (University of California at Los Angeles) for tRNAUGASer(U47:6), D. Patel for providing coordinates before publication and J. Anderson for advice, as well as S. Wolin, The Friday Seminar, R. Crouch and members of the Laboratory of Molecular Growth Regulation for discussion, comments or both, and A. Hinnebusch for suggesting the hypomorph and for comments on the manuscript. This work was supported by the Intramural Research Program of the National Institute of Child Health and Human Development, US National Institutes of Health. R.J.M. serves as an Officer in the US Public Health Service.

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  1. Ying Huang and Mark A Bayfield: These authors contributed equally to this work.

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  1. Laboratory of Molecular Growth Regulation, National Institute of Child Health and Human Development, US National Institutes of Health, 31 Center Dr., Rm. 2A25, Bethesda, 20892, Maryland, USA

    Ying Huang, Mark A Bayfield, Robert V Intine & Richard J Maraia

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Y.H., M.A.B. and R.V.I. designed and performed experiments. R.J.M. designed experiments and wrote the manuscript

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Correspondence to Richard J Maraia.

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Huang, Y., Bayfield, M., Intine, R. et al. Separate RNA-binding surfaces on the multifunctional La protein mediate distinguishable activities in tRNA maturation. Nat Struct Mol Biol 13, 611–618 (2006). https://doi.org/10.1038/nsmb1110

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