The terminal balls characteristic of eukaryotic rRNA transcription units in chromatin spreads are rRNA processing complexes

• Published in: Genes & development 1993-08, Vol.7 (8), p.1609-1619
• Main Authors: MOUGEY, E. B, O'REILLY, M, OSHEIM, Y, MILLER, O. L, BEYER, A, SOLLNER-WEBB, B
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Biological and medical sciences; Chromatin - ultrastructure; Conserved Sequence; DNA, Ribosomal - ultrastructure; Fundamental and applied biological sciences. Psychology; Microscopy, Electron; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Mutagenesis, Insertional; Oocytes - ultrastructure; Repetitive Sequences, Nucleic Acid; RNA Precursors - ultrastructure; RNA Processing, Post-Transcriptional; RNA, Ribosomal - ultrastructure; Sequence Analysis, RNA; Transcription, Genetic; Transcription. Transcription factor. Splicing. Rna processing; Xenopus laevis; Xenopus laevis - genetics
• ISSN: 0890-9369; 1549-5477
• DOI: 10.1101/gad.7.8.1609

When spread chromatin is visualized by electron microscopy, active rRNA genes have a characteristic Christmas tree appearance: From a DNA "trunk" extend closely packed "branches" of nascent transcripts whose ends are decorated with terminal "balls." These terminal balls have been known for more than two decades, are shown in most biology textbooks, and are reported in hundreds of papers, yet their nature has remained elusive. Here, we show that a rRNA-processing signal in the 5'-external transcribed spacer (ETS) of the Xenopus laevis ribosomal primary transcript forms a large, processing-related complex with factors of the Xenopus oocyte, analogous to 5' ETS processing complexes found in other vertebrate cell types. Using mutant rRNA genes, we find that the same rRNA residues are required for this biochemically defined complex formation and for terminal ball formation, analyzed electron microscopically after injection of these cloned genes into Xenopus oocytes. This, plus other presented evidence, implies that rRNA terminal balls in Xenopus, and by inference, also in the multitude of other species where they have been observed, are the ultrastructural visualization of an evolutionarily conserved 5' ETS processing complex that forms on the nascent rRNA.