PMID: 6164981Feb 25, 1981Paper

Purification and characterization of a simple ribonucleoprotein particle containing small nucleoplasmic RNAs (snRNP) as a subset of RNP containing heterogenous nuclear RNA (hnRNP) from HeLa cells

Nucleic Acids Research
C BrunelJ P Liautard

Abstract

A ribonucleoprotein complex whose RNA complement consists exclusively of small nuclear RNA species (snRNA) has been purified from particles containing heterogenous nuclear RNA (hnRNP) from HeLa cells. This was accomplished by taking advantage of their ability to band at a density of about 1.43 g/cm3 in plain cesium chloride as well as in cesium chloride gradients containing 0.5% sarkosyl without prior aldehyde fixation. After these two steps of equilibrium density centrifugation, these snRNPs were still largely contaminated by free proteins (and especially phosphoproteins). A final step of purification by velocity sedimentation in a sucrose gradient containing 0.5 M cesium chloride and 0.5% sarkosyl was efficient in completely eliminating all free proteins. U1, U2, U4, U5 and U6 species according to the nomenclature of Lerner et al. (Nature, (1980) 283, 220-224) were found in these purified snRNPs, while a significant part of U6 and a small amount of U2 were found in the bottom fraction. 5S species behaved entirely as free RNA and is presumably a contaminant of cytoplasmic origin. Electrophoresis of proteins from snRNP labeled in vivo with (35S) methionine, revealed four bands with migrations corresponding to molecular weights ...Continue Reading

References

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Citations

Sep 1, 1992·Molecular Biology Reports·V SumpterR Lührmann
Jun 1, 1990·Molecular and Biochemical Parasitology·S Michaeli, N Agabian
Nov 30, 1990·Biochimica Et Biophysica Acta·R LührmannM Bach
Mar 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·I V Economidis, T Pederson
Jul 1, 1983·The Journal of Cell Biology·D J ForbesM W Kirschner
Jun 1, 1987·Clinical Rheumatology·D Williams
Jun 5, 1986·Journal of Molecular Biology·M N Lelay-TahaP Jeanteur
May 15, 1985·Analytical Biochemistry·W H SchrierT B Okarma
May 15, 1985·Analytical Biochemistry·T B OkarmaR Feinbaum
Jun 15, 1982·Biochemical and Biophysical Research Communications·C AssensP Jeanteur
Jan 29, 1982·Biochemical and Biophysical Research Communications·J Sri-WidadaC Brunel
Feb 1, 1990·Molecular Biology Reports·K Scherrer
Feb 20, 1986·Journal of Molecular Biology·C Martins de SaH P Schmid

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