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  Cryoelectron microscopy and cryoelectron tomography of the nuclear pre-mRNA processing machine

Medalia, O., Typke, D., Hegerl, R., Angenitzki, M., Sperling, J., & Sperling, R. (2002). Cryoelectron microscopy and cryoelectron tomography of the nuclear pre-mRNA processing machine. Journal of Structural Biology, 138(1-2), 74-84.

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Genre: Journal Article
Alternative Title : J. Struct. Biol.

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 Creators:
Medalia, O.1, Author              
Typke, D.2, Author              
Hegerl, R.2, Author              
Angenitzki, M., Author
Sperling, J., Author
Sperling, R., Author
Affiliations:
1External Organizations, ou_persistent22              
2Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565142              

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Free keywords: InRNP complexes; cryoelectron microscopy; charged lipid monolayer; cryoelectron tomography; image restoration; three- dimensional image reconstruction; pre-mRNA processing
 Abstract: Large nuclear ribonucleoprotein particles, which can be viewed as the naturally assembled precursor messenger RNA (pre-mRNA) processing machine, were analyzed in frozen-hydrated preparations by cryoelectron microscopy. A general and reproducible strategy for preparing ice-embedded large nuclear ribonucleoprotein (lnRNP) particles at sufficiently high concentration was developed. Taking advantage of their negatively charged components, the lnRNP particles are adsorbed and thus concentrated on a positively charged lipid monolayer while preserving their native structure. Using this approach we carried out cryoelectron tomography and three-dimensional image reconstruction of individual InRNP particles. The study revealed a structure similar to that of negatively stained particles studied previously, yet with additional features. The small additional domain visualized in negative stain appeared to be larger in the ice preparations. In addition, using image restoration from focus series of ice-embedded InRNP particles, new features such as holes within the subunits were visualized in two dimensions, and it was shown that the subunits are interconnected via a fiber, very likely formed by the pre-mRNA. This finding supports the model that each subunit represents a spliceosome that splices out the intron wound around it. (C) 2002 Elsevier Science (USA). All rights reserved.

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Language(s): eng - English
 Dates: 2002-04
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 41768
ISI: 000177978800008
 Degree: -

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Title: Journal of Structural Biology
  Alternative Title : J. Struct. Biol.
Source Genre: Journal
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Publ. Info: -
Pages: - Volume / Issue: 138 (1-2) Sequence Number: - Start / End Page: 74 - 84 Identifier: ISSN: 1047-8477