Transcriptional termination in the Balbiani ring 1 gene is closely coupled to 3'-end formation and excision of the 3'-terminal intron

We have analyzed transcription termination, 3'-end formation, and excision of the 3'-terminal intron in vivo in the Balbiani ring 1 (BR1) gene and its pre-mRNA. We show that full-length RNA transcripts are evenly spaced on the gene from a position 300 bp upstream to a region 500-700 bp dow...

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Bibliographic Details
Published in:Genes & development 1998-09, Vol.12 (17), p.2759-2769
Main Authors: Baurén, G, Belikov, S, Wieslander, L
Format: Article
Language:English
Subjects:
Animals
Chironomidae - genetics
Chironomidae - metabolism
Genes, Insect
Insect Proteins - biosynthesis
Insect Proteins - genetics
Introns
Polymerase Chain Reaction
Regulatory Sequences, Nucleic Acid
Research Paper
RNA Precursors - biosynthesis
RNA Precursors - genetics
Salivary Glands - metabolism
Sequence Deletion
Transcription, Genetic
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Summary:We have analyzed transcription termination, 3'-end formation, and excision of the 3'-terminal intron in vivo in the Balbiani ring 1 (BR1) gene and its pre-mRNA. We show that full-length RNA transcripts are evenly spaced on the gene from a position 300 bp upstream to a region 500-700 bp downstream of the polyadenylation sequence. Very few full-length transcripts and no short, cleaved, nascent transcripts could be observed downstream of this region. Pre-mRNA with 10-20 adenylate residues accumulates at the active gene and then rapidly leaves from the gene locus. Only polyadenylated pre-mRNAs could be detected in the nucleoplasm. Our results are consistent with the hypothesis that transcription termination occurs in a narrow region for the majority of transcripts, simultaneous with 3'-end formation. Excision of the 3'-terminal intron occurs before 3'-end formation in about 5% of the nascent transcripts. When transcription terminates, 3' cleavage takes place and 10-20 adenylate residues are added, the 3'-terminal intron is excised from additionally about 75% of the pre-mRNA at the gene locus. Our data support a close temporal and spatial coupling of transcription termination and the cleavage and initial polyadenylation of 3'-end formation. Excision of the 3'-terminal intron is highly stimulated as the cleavage/polyadenylation complex assembles and 3'-end formation is initiated.
ISSN:0890-9369
1549-5477
DOI:10.1101/gad.12.17.2759