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Lsm proteins bind and stabilize RNAs containing 5′ poly(A) tracts

Nature Structural & Molecular Biology volume 14, pages 824–831 (2007)Cite this article

Abstract

Many orthopoxvirus messenger RNAs have an unusual nontemplated poly(A) tract of 5 to 40 residues at the 5′ end. The precise function of this feature is unknown. Here we show that 5′ poly(A) tracts are able to repress RNA decay by inhibiting 3′-to-5′ exonucleases as well as decapping of RNA substrates. UV cross-linking analysis demonstrated that the Lsm complex associates with the 5′ poly(A) tract. Furthermore, recombinant Lsm1–7 complex specifically binds 5′ poly(A) tracts 10 to 21 nucleotides in length, consistent with the length of 5′ poly(A) required for stabilization. Knockdown of Lsm1 abrogates RNA stabilization by the 5′ poly(A) tract. We propose that the Lsm complex simultaneously binds the 3′ and 5′ ends of these unusual messenger RNAs and thereby prevents 3′-to-5′ decay. The implications of this phenomenon for cellular mRNA decay are discussed.

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Figure 1: A 5′ adenosine tract of at least 10 residues inhibits RNA decay in cells.
Figure 2: 5′ adenosine tracts block both 3′-to-5′ decay and decapping in vitro.
Figure 3: PABP and Lsm proteins associate specifically with the 5′ adenosine tract.
Figure 4: Purified recombinant Lsm complex binds specifically to RNAs bearing a 5′ adenosine tract.
Figure 5: Purified recombinant Lsm complex requires a 5′ adenosine tract of at least 10 nt.
Figure 6: Knockdown of Lsm1 blocks the ability of a 5′ adenosine tract to stabilize RNAs.
Figure 7: Lsm proteins may circularize RNAs with 5′ adenosine tracts, there by inhibiting both 3′-to-5′ and 5′-to-3′ decay.

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Acknowledgements

This work was supported by a grant from the US National Institutes of Health (GM72481) to J.W. B.Z. contributed to the paper while receiving Swiss National Foundation funds in the laboratory of C.K. We thank E.K.L. Chan (University of Florida) for supplying the Lsm4-specific antibodies, K. Sokoloski (Colorado State University) for the sequence of hamster Lsm1 and D. Pickup (Duke University) for helpful discussions.

Author information

Author notes

  1. Naomi Bergman & Karen C M Moraes

    Present address: Present addresses: New York University School of Medicine, Department of Microbiology, New York, New York 10016, USA (N.B.) and Universidade do Vale do Paraíba, IP&D, Av. Shishima Hifumi, 2911, Urbanova, CEP 12244-000, São José dos Campos, SP, Brazil (K.C.M.M.).,

  2. Naomi Bergman and Karen C M Moraes: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology, Immunology & Pathology, Colorado State University, Fort Collins, 80525, Colorado, USA

    Naomi Bergman, Karen C M Moraes, John R Anderson, Carol J Wilusz & Jeffrey Wilusz

  2. Vinca Institute of Nuclear Sciences, Laboratory of Molecular Biology and Endocrinology, Mihaila Petrovica Alasa 12-14, Belgrade, PO Box 522-090, Serbia

    Bozidarka Zaric

  3. Paul Scherrer Institut, Biomolecular Research, Villigen, CH5232, Switzerland

    Christian Kambach

  4. Department of Microbiology, New York University School of Medicine, New York, 10016, New York, USA

    Robert J Schneider

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Contributions

K.C.M.M. and N.B. performed most of the experiments and contributed to their analysis and design. N.B. initially discovered the novel role and properties of the Lsm complex, and also constructed the Lsm1-knockdown cells together with R.J.S. B.Z. and C.K. purified and reconstituted the Lsm complexes. C.J.W. wrote a large portion of the paper and contributed to experimental design and analysis. J.W. was conceived the project, was involved in design and analysis of experiments, and performed some of the electroporation studies with assistance from J.R.A.

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Correspondence to Jeffrey Wilusz.

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Bergman, N., Moraes, K., Anderson, J. et al. Lsm proteins bind and stabilize RNAs containing 5′ poly(A) tracts. Nat Struct Mol Biol 14, 824–831 (2007). https://doi.org/10.1038/nsmb1287

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