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Mapping structural differences between 30S ribosomal subunit assembly intermediates

Nature Structural & Molecular Biology volume 11, pages 179–186 (2004)Cite this article

Abstract

Under appropriate conditions, functional Escherichia coli 30S ribosomal subunits assemble in vitro from purified components. However, at low temperatures, assembly stalls, producing an intermediate (RI) that sediments at 21S and is composed of 16S ribosomal RNA (rRNA) and a subset of ribosomal proteins (r-proteins). Incubation of RI at elevated temperatures produces a particle, RI*, of similar composition but different sedimentation coefficient (26S). Once formed, RI* rapidly associates with the remaining r-proteins to produce mature 30S subunits. To understand the nature of this transition from RI to RI*, changes in the reactivity of 16S rRNA between these two states were monitored by chemical modification and primer extension analysis. Evaluation of this data using structural and biochemical information reveals that many changes are r-protein–dependent and some are clustered in functional regions, suggesting that this transition is an important step in functional 30S subunit formation.

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Figure 1: In vitro assembly of 30S ribosomal subunits.
Figure 2: 5′ domain nucleotides with altered reactivity between RI and RI*.
Figure 3: Central domain nucleotides with altered reactivity between RI and RI*.
Figure 4: 3′ domain nucleotides with altered reactivity between RI and RI*.
Figure 5: Summary of nucleotide changes between RI and RI*.
Figure 6: Sites of decreased reactivity in RI* relative to RI.
Figure 7: Sites of enhanced reactivity in RI* relative to RI.

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Acknowledgements

We thank R. Green, I. Jagannathan and J. Maki for critical reading of the manuscript. Additional thanks to S. Stagg and J. Hoy for assistance with figures. This work was funded by a grant from the US National Institutes of Health (to G.M.C.).

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  1. Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, 50011, Iowa, USA

    Kristi L Holmes & Gloria M Culver

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Correspondence to Gloria M Culver.

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Holmes, K., Culver, G. Mapping structural differences between 30S ribosomal subunit assembly intermediates. Nat Struct Mol Biol 11, 179–186 (2004). https://doi.org/10.1038/nsmb719

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