Affinity purification of Trypanosoma brucei small nuclear ribonucleoproteins reveals common and specific protein components.
We have developed a procedure for the affinity purification of small nuclear ribonucleoproteins (snRNPs) of Trypanosoma brucei (U2 and U4/U6 snRNPs), which are essential for trans splicing. Each of these snRNPs can be specifically and efficiently selected from T. brucei extracts through biotinylated antisense 2'-O-methylated RNA oligonucleotides immobilized on streptavidin-agarose. Protein analysis revealed a set of five low molecular weight polypeptides common to the U2 and U4/U6 snRNPs and the spliced leader RNP. In addition, several U2 and U4/U6 snRNP-specific protein components were identified. Using monoclonal antibodies against human snRNP proteins, we could not detect any significant cross-reaction with the trypanosomal U2 snRNP proteins. Thus, the trypanosomal snRNPs exhibit principal differences from the higher eukaryotic snRNPs not only in their RNA but also in their protein components.
Palfi
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Max-Planck-Institut für Molekulare Genetik, Otto-Warburg-Laboratorium, Federal Republic of Germany.
Günzl
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D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D003429:Cross Reactions; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016376:Oligonucleotides, Antisense; D012326:RNA Splicing; D012342:RNA, Small Nuclear / Q000235:genetics / Q000302:isolation & purification*; D012261:Ribonucleoproteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism*; D017411:Ribonucleoproteins, Small Nuclear; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D016376:Oligonucleotides, Antisense; D012342:RNA, Small Nuclear; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear