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          Institute: MPI für biophysikalische Chemie     Collection: Zelluläre Biochemie (Prof. Reinhard Lührmann)     Display Documents

ID: 32817.0, MPI für biophysikalische Chemie / Zelluläre Biochemie (Prof. Reinhard Lührmann)
Direct probing of RNA structure and RNA-protein interactions in purifies HeLa cells and yeast spliceosomal U4/U6.U5 tri-snRNP particles
Authors:Mougin, A.; Gottschalk, A.; Fabrizio, P.; Luehrmann, R.; Branlant, C.
Date of Publication (YYYY-MM-DD):2002
Title of Journal:Journal of Biological Chemistry
Issue / Number:5
Start Page:631
End Page:649
Review Status:not specified
Audience:Not Specified
Abstract / Description:The U4/U6.U5 tri-snRNP is a key component of spliceosomes. By using chemical reagents and
RNases, we performed the first extensive experimental analysis of the structure and accessibility of
U4 and U6 snRNAs in tri-snRNPs. These were purified from HeLa cell nuclear extract and
Saccharomyces cerevisiae cellular extract. U5 accessibility was also investigated. For both species,
data demonstrate the formation of the U4/U6 Y-shaped structure. In the human tri-snRNP and
U4/U6 snRNP, U6 forms the long range interaction, that was previously proposed to be
responsible for dissociation of the deproteinized U4/U6 duplex. In both yeast and human
tri-snRNPs, U5 is more protected than U4 and U6, suggesting that the U5 snRNP-specific protein
complex and other components of the tri-snRNP wrapped the 5' stem-loop of U5. Loop I of U5 is
partially accessible, and chemical modifications of loop I were identical in yeast and human
tri-snRNPs. This reflects a strong conservation of the interactions of proteins with the functional
loop I. Only some parts of the U4/U6 Y-shaped motif (the 5' stem-loop of U4 and helix II) are
protected. Due to difference of protein composition of yeast and human tri-snRNP, the U6 segment
linking the 5' stem-loop to the Y-shaped structure and the U4 central single-stranded segment are
more accessible in the yeast than in the human tri-snRNP, especially, the phylogenetically conserved
ACAGAG sequence of U6. Data are discussed taking into account knowledge on RNA and
protein components of yeast and human snRNPs and their involvement in splicesome assembly.
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für biophysikalische Chemie/Abt. Reinhard Lührmann / 100
External Affiliations:UMR 7567 CNRS-UHP Nancy I, Maturation des ARN et Enzymologie Moléculaire, Université H. Poincaré, B.P. 239, 54506, Vandoeuvre-les Nancy Cédex, France
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