Abstract
Publisher Summary This chapter describes methods for the isolation of these ribonucleoprotein (RNP) particles from eukaryotic cells and discusses some properties of heterogeneous nuclear RNA (hnRNA) particles isolated from HeLa cell nuclei. RNP particles containing hnRNA have been identified both cytologically and biochemically in eukaryotic cell nuclei. The method developed for isolation of HeLa hnRNP particles involves the isolation of nuclei followed by their mechanical disruption and purification of hnRNP particles by sucrose gradient centrifugation. The method of disrupting HeLa cell nuclei permits the high recovery of heterogeneous nuclear RNA in the presence of large amounts of nucleohistone. An understanding of the specificity of RNA-protein interactions in native hnRNP particles is impeded by the nucleotide sequence complexity of hnRNA and by the analytical complexity of the proteins. The specific association of protein with poly (A)-rich hnRNA sequences is demonstrated.
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