Immunological characterization and intracellular localization of trans-spliceosomal small nuclear ribonucleoproteins in Trypanosoma brucei.
Polyclonal antibodies were raised against purified protein components of the U2 small nuclear ribonucleoprotein (snRNP) from Trypanosoma brucei. Through immunoblot and immunoprecipitation analyses three antisera were characterized that reacted specifically with U2 snRNP proteins of molecular weights 40,000 (anti-40K) and 16,500 (anti-16.5K), and with each of four proteins of molecular weights 14,000, 12,500, 10,000, and 8,500 (anti-CP). Anti-40K antibodies specifically immunoprecipitated the U2 snRNP from trypanosomal extracts, whereas anti-CP antibodies recognized several snRNPs, including the SL RNP and the U2 and U4/U6 snRNPs; in addition, minor RNAs were detected, suggesting that a family of snRNPs with common or related protein components exists in trypanosomes. None of these antibodies cross-reacted significantly with total mammalian snRNP proteins, indicating that the trypanosomal snRNP proteins are immunologically distinct from their mammalian counterparts. Using immunofluorescence microscopy, the snRNP proteins exhibited a differential cellular distribution. Whereas the 40-kDa protein is localized exclusively in the nucleus, with the nucleolus being excluded, a fraction of the common proteins also resides in the cytoplasm.
Palfi
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Max-Planck-Institut für Molekulare Genetik, Otto-Warburg-Laboratorium, Berlin, (Dahlem), Federal Republic of Germany.
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D000818:Animals; D000906:Antibodies / Q000276:immunology; D015153:Blotting, Western; D002467:Cell Nucleus / Q000737:chemistry; D003429:Cross Reactions; D003593:Cytoplasm / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D011233:Precipitin Tests; D017411:Ribonucleoproteins, Small Nuclear / Q000032:analysis* / Q000276:immunology; D017381:Spliceosomes / Q000737:chemistry*; D014346:Trypanosoma brucei brucei / Q000737:chemistry*
D000906:Antibodies; D017411:Ribonucleoproteins, Small Nuclear