RNA-Protein Interaction Protocols

Volume 118 of the series Methods in Molecular Biology™ pp 299-308

Preparation of Heterogeneous Nuclear Ribonucleoprotein Complexes

  • Maurice S. SwansonAffiliated withDepartment of Molecular Genetics and Microbiology and Centers for Gene Therapy and Mammalian Genetics, University of Florida College of Medicine
  • , Gideon DreyfussAffiliated withHoward Hughes Medical Institute, Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine

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Proper formation of different types of ribonucleoprotein (RNP)-RNA complexes is critical for the correct expression of genetic information at the posttranscriptional level. In the nucleus, RNA polymerase II transcripts bind to an abundant class of nuclear pre-mRNA/mRNA-binding proteins, the heterogeneous nuclear ribonucleoproteins (hnRNPs), to form hnRNP complexes (1,2). The molecular architecture of these complexes must be exceedingly dynamic because each step in the pre-mRNA processing pathway alters the structure of the underlying RNA substrate. Therefore, hnRNP complexes within the cell, although probably unique for each transcript, are not uniform structures, such as a ribosomal subunit or a particular small nuclear ribonucleoprotein particle (snRNP), but are composed of a heterogeneous mixture of protein-RNA structures.