Crystallizing the 6S and 8S spliceosomal assembly intermediates : a complex project

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2015
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Pelz, Jann-Patrick
Schindelin, Hermann
van Pee, Katharina
Kuper, Jochen
Kisker, Caroline
Fischer, Utz
Grimm, Clemens
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Acta Crystallographica Section D : Biological Crystallography. 2015, 71(10), pp. 2040-2053. ISSN 0907-4449. eISSN 1399-0047. Available under: doi: 10.1107/S1399004715014832
Zusammenfassung

The small nuclear ribonucleoproteins (snRNPs) U1, U2, U4/6 and U5 are major constituents of the pre-mRNA processing spliceosome. They contain a common RNP core that is formed by the ordered binding of Sm proteins onto the single-stranded Sm site of the snRNA. Although spontaneous in vitro, assembly of the Sm core requires assistance from the PRMT5 and SMN complexes in vivo. To gain insight into the key steps of the assembly process, the crystal structures of two assembly intermediates of U snRNPs termed the 6S and 8S complexes have recently been reported. These multimeric protein complexes could only be crystallized after the application of various rescue strategies. The developed strategy leading to the crystallization and solution of the 8S crystal structure was subsequently used to guide a combination of rational crystal-contact optimization with surface-entropy reduction of crystals of the related 6S complex. Conversely, the resulting high-resolution 6S crystal structure was used during the restrained refinement of the 8S crystal structure.

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Fachgebiet (DDC)
570 Biowissenschaften, Biologie
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U snRNP assembly, spliceosomal assembly, contact engineering, surface-entropy reduction, SMN, Sm proteins, pICln
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ISO 690PELZ, Jann-Patrick, Hermann SCHINDELIN, Katharina VAN PEE, Jochen KUPER, Caroline KISKER, Kay DIEDERICHS, Utz FISCHER, Clemens GRIMM, 2015. Crystallizing the 6S and 8S spliceosomal assembly intermediates : a complex project. In: Acta Crystallographica Section D : Biological Crystallography. 2015, 71(10), pp. 2040-2053. ISSN 0907-4449. eISSN 1399-0047. Available under: doi: 10.1107/S1399004715014832
BibTex
@article{Pelz2015Cryst-32539,
  year={2015},
  doi={10.1107/S1399004715014832},
  title={Crystallizing the 6S and 8S spliceosomal assembly intermediates : a complex project},
  number={10},
  volume={71},
  issn={0907-4449},
  journal={Acta Crystallographica Section D : Biological Crystallography},
  pages={2040--2053},
  author={Pelz, Jann-Patrick and Schindelin, Hermann and van Pee, Katharina and Kuper, Jochen and Kisker, Caroline and Diederichs, Kay and Fischer, Utz and Grimm, Clemens}
}
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