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Phosphorothioate oligonucleotides bind in a non sequence‐specific manner to the nucleolar protein C23/nucleolin
Author(s) -
Weidner Douglas A.,
Valdez Benigno C.,
Henning Dale,
Greenberg Scott,
Busch Harris
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)00517-d
Subject(s) - nucleolin , oligonucleotide , microbiology and biotechnology , immunoprecipitation , binding protein , nuclear protein , biology , fusion protein , biochemistry , rna , chemistry , cytoplasm , recombinant dna , dna , nucleolus , transcription factor , gene
To design optimal strategies for intracellular delivery of antisense phosphorothioate oligonucleotides, it may be useful to understand their interaction with cellular macromolecules. Nuclear extracts from LOX amelanotic myeloma cells were studied for protein binding to phosphorothioate oligonucleotides using a Southwestern protocol. Multiple nuclear proteins bound to the phosphorothioate oligonucleotides but no detectable protein binding was found to phosphodiester oligonucleotides. The protein with the strongest binding signals was shown by immunoprecipitation to be nucleolar C23/nucleolin, a 110 kDa protein. With glutathione S‐transferase/nucleolin fusion protein constructs, the region of nucleolin containing the RNA recognition motifs had binding activity to phosphorothioate oligonucleotides.