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PMID: 2356121 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Permeable trypanosome cells as a model system for transcription and trans-splicing.

Nucleic acids research ·Vol. 18 ·No. 11 ·1990-06-11 ·Pages 3319-26

Ullu E, Tschudi C

Abstract

We have established conditions for Trypanosoma brucei permeable cells to study transcription and trans-splicing. We found that the concentration of monovalent and, to a lesser extent, divalent ions plays a critical role for the expression of a number of different genes. Most remarkably, the synthesis of the spliced leader (SL) RNA was optimal at 20 mM KCl, whereas higher potassium concentrations were inhibitory. In addition, MgCl2 concentrations above 3 mM led to the accumulation of a 3' end shortened SL RNA species, which has been previously reported not to participate in trans-splicing. Using conditions optimal for the synthesis of the SL RNA, we observed accurate trans-splicing of newly-synthesized alpha-tubulin RNA. Moreover, we detected the SL intron both joined to high molecular weight RNAs in the form of branched Y-structures and as a free linear molecule, which rapidly turned over. Furthermore, ionic concentrations that inhibit the synthesis of the SL RNA produced exclusively unspliced alpha-tubulin RNA, thus demonstrating that transcription and trans-splicing can be uncoupled.

MeSH Terms
Amanitins/pharmacology Animals Base Sequence Cells, Cultured Exons Gene Expression Regulation Introns Magnesium Chloride/pharmacology Molecular Sequence Data Permeability Potassium Chloride/pharmacology RNA Splicing RNA, Messenger/biosynthesis,genetics Transcription, Genetic Trypanosoma brucei brucei/genetics
Chemicals
Amanitins RNA, Messenger Magnesium Chloride Potassium Chloride
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Ullu E
Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06510.
Tschudi C
References (32)
32 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/18.11.3319
Published
1990-06-11
Pages
3319-26
Language
English
Region
England
NLM ID
0411011
PMCID
PMC330939
Subset
IM
Grants
NIAID NIH HHS · AI08614 · United States
External Links
PubMed source DOI: 10.1093/nar/18.11.3319 PMC Full Text View journal details
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