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  Three-dimensional image reconstruction of large nuclear rnp (lnrnp) particles by automated electron tomography

Sperling, R., Koster, A. J., Melamedbessudo, C., Rubinstein, A., Angenitzki, M., Berkovitchyellin, Z., et al. (1997). Three-dimensional image reconstruction of large nuclear rnp (lnrnp) particles by automated electron tomography. Journal of Molecular Biology, 267(3), 570-583.

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Sperling, R., Autor
Koster, A. J.1, Autor           
Melamedbessudo, C., Autor
Rubinstein, A., Autor
Angenitzki, M., Autor
Berkovitchyellin, Z., Autor
Sperling, J., Autor
Affiliations:
1Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565142              

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Schlagwörter: Pre-mrna splicing; Lnrnp particles; Model; Electron microscopy.; Pre-messenger-rna; Splice-site selection; 3-dimensional structure; Ribonucleoprotein-particles; Microscope tomography; Inrnp particles; Visualization; Micrographs; Assemblies.; Molecular biology & genetics.
 Zusammenfassung: Nuclear RNA transcripts of split genes and their splicing products, as well as the general population of nuclear polyadenylated RNA are packaged in multi-component large nuclear ribonucleoprotein (lnRNP) particles. These lnRNP particles, which sediment at the 200 S region in sucrose gradients, contain all U small nuclear RNPs required for precursor messenger RNA (pre-mRNA) splicing and several protein splicing factors, including U2AF and the SR proteins. Electron microscopy of lnRNP particles revealed a large compact structure of 50 nm in diameter. In this study we employed automated computed tomography from electron micrographs for the three-dimensional (3D) image reconstruction of individual lnRNP particles isolated from mammalian cells nuclei and negatively stained. For each particle, a tilt series of 71 images was collected by direct digital recording of the images on a CCD camera attached to a computer controlled TEM facility. The 3D image was reconstructed according to the back projection principle. For rendering, real time display and comparison of the reconstructed particles, interactive computer graphics was employed. The reconstructed 3D images show a compact structure composed of four major subunits connected to each other. Comparison of the reconstructed lnRNP particles revealed morphological similarity of the individual particles, as well as similarity among the sub-structures. Based on these observations we propose a model for the packaging of nuclear pre-mRNAs in lnRNP particles where each sub-structure represents a functional unit. This model is compatible with the requirements for alternative splicing in multi-intronic pre-mRNAs, and with the fact that the splicing of multi-intronic pre-mRNAs does not occur in a sequential manner. (C) 1997 Academic Press Limited. [References: 49]

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 Datum: 1997-04-04
 Publikationsstatus: Erschienen
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 Identifikatoren: eDoc: 318597
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Titel: Journal of Molecular Biology
Genre der Quelle: Zeitschrift
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Ort, Verlag, Ausgabe: -
Seiten: - Band / Heft: 267 (3) Artikelnummer: - Start- / Endseite: 570 - 583 Identifikator: -