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  Reconstitution of the complete pathway of ITS2 processing at the pre-ribosome

Fromm, L., Falk, S., Flemming, D., Schuller, J. M., Thoms, M., Conti, E., et al. (2017). Reconstitution of the complete pathway of ITS2 processing at the pre-ribosome. Nature Communications, 8: 1787. doi:10.1038/s41467-017-01786-9.

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Fromm, Lisa1, Author
Falk, Sebastian2, Author              
Flemming, Dirk1, Author
Schuller, Jan Michael2, Author              
Thoms, Matthias1, Author
Conti, Elena2, Author              
Hurt, Ed1, Author
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1external, ou_persistent22              
2Conti, Elena / Structural Cell Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565144              

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 Abstract: Removal of internal transcribed spacer 2 (ITS2) from pre-ribosomal RNA is essential to make functional ribosomes. This complicated processing reaction begins with a single endonucleolytic cleavage followed by exonucleolytic trimming at both new cleavage sites to generate mature 5.8S and 25S rRNA. We reconstituted the 7S -> 5.8S processing branch within ITS2 using purified exosome and its nuclear cofactors. We find that both Rrp44's ribonuclease activities are required for initial RNA shortening followed by hand over to the exonuclease Rrp6. During the in vitro reaction, ITS2-associated factors dissociate and the underlying 'foot' structure of the pre-60S particle is dismantled. 7S pre-rRNA processing is independent of 5S RNP rotation, but 26S -> 25S trimming is a precondition for subsequent 7S -> 5.8S processing. To complete the in vitro assay, we reconstituted the entire cycle of ITS2 removal with a total of 18 purified factors, catalysed by the integrated activities of the two participating RNA-processing machines, the Las1 complex and nuclear exosome.

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 Dates: 2017-11-27
 Publication Status: Published online
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Title: Nature Communications
  Abbreviation : Nat. Commun.
Source Genre: Journal
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Publ. Info: London : Nature Publishing Group
Pages: - Volume / Issue: 8 Sequence Number: 1787 Start / End Page: - Identifier: ISSN: 2041-1723
CoNE: https://pure.mpg.de/cone/journals/resource/2041-1723