In vitro splicing of the ribosomal RNA precursor in isolated nucleoli from Tetrahymena

The macronuclear rRNA genes of Tetrahymena thermophila contain an 0.4 kb intervening sequence in the 26S rRNA coding region. The sequence is represented within the primary transcription product. We demonstrate in this paper that the enzyme activities necessary for the endonucleolytic cleavage as wel...

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Published in:Nucleic acids research 1980-12, Vol.8 (23), p.5551-5566
Main Authors: Carin, M., Jensen, B.F., Jentsch, K.D., Leer, J.C., Nielsen, O.F., Westergaard, O.
Format: Article
Language:English
Subjects:
Animals
Cell Nucleolus - metabolism
DNA - genetics
Genes
In Vitro Techniques
Nucleic Acid Precursors - metabolism
RNA, Ribosomal - metabolism
Tetrahymena - metabolism
Tetrahymena thermophila
Transcription, Genetic
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container_end_page 5566
container_issue 23
container_start_page 5551
container_title Nucleic acids research
container_volume 8
creator Carin, M.
Jensen, B.F.
Jentsch, K.D.
Leer, J.C.
Nielsen, O.F.
Westergaard, O.
description The macronuclear rRNA genes of Tetrahymena thermophila contain an 0.4 kb intervening sequence in the 26S rRNA coding region. The sequence is represented within the primary transcription product. We demonstrate in this paper that the enzyme activities necessary for the endonucleolytic cleavage as well as for the ligation of the transcript are associated with the isolated nucleoli. Both of these processes occur posttranscriptionally. The intervening sequence is excised as an unique molecule, which is stable in vitro. About 50% of the in vitro synthesized RNA is processed. Faithful in vitro transcription occurs in the presence of the divalent ions Mg2+, Mn2+ and Co2+ while processing takes place only in the presence of Mg2+. The absolute requirement for Mg2+ in the excision reaction enables us to synthesize labelled pre-rRNA in the presence of Mn2+ or Co2+. The synthesized RNA can be used as a substrate in studies of the processing enzymes in vitro.
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subjects Animals
Cell Nucleolus - metabolism
DNA - genetics
Genes
In Vitro Techniques
Nucleic Acid Precursors - metabolism
RNA, Ribosomal - metabolism
Tetrahymena - metabolism
Tetrahymena thermophila
Transcription, Genetic
title In vitro splicing of the ribosomal RNA precursor in isolated nucleoli from Tetrahymena
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