Immunoelectron microscope visualization of nuclear ribonucleoprotein antigens within spread transcription complexes

Abstract

The ultrastructural distribution of nuclear ribonucleoproteins (RNP) within spread active chromatin has been investigated using specific anti-RNP antibodies. Monoclonal antibodies directed against the core proteins of heterogeneous nuclear (hn)RNP or against small nuclear (sn)RNP have been incubated directly with lysed mouse or Drosophila tissue culture cells and the bound antibodies visualized by means of a protein A-colloidal gold complex. The hnRNP core proteins have been localized on growing RNP fibrils within non-nucleolar transcription complexes. Anti-snRNP antibodies, directed either against the Sm-antigen (common for nucleoplasmic snRNP species containing U1, U2, U4, U5, and U6 RNAs) or against U1-snRNP, were bound by two morphological types of RNP structures. Within areas of chromatin that do not completely disperse, labeling was observed on RNP-fibril gradient type structures or on groups of fibrogranular material. In the well dispersed regions containing individual nonribosomal transcription complexes, snRNP antigens were associated with growing RNP fibrils. Our results provide direct evidence for association of some U-snRNP species (including U1-snRNP) with extranucleolar RNA as early as during transcription elongation. In addition, the presence of core hnRNP proteins on the same type of nascent RNA transcripts has been confirmed.