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Nonsense suppression inDictyostelium discoideum

✍ Scribed by Dingermann, Theodor ;Reindl, Norbert ;Brechner, Thomas ;Werner, Herbert ;Nerke, Käthe

Book ID
102820767
Publisher
John Wiley and Sons
Year
1990
Tongue
English
Weight
836 KB
Volume
11
Category
Article
ISSN
0192-253X

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✦ Synopsis

We describe the generation of Dictyosteliurn discoideurn cell lines that carry different suppressor tRNA genes. These genes were constructed by primer-directed mutagenesis changing a tRNATrP(CCA) gene from D. discoideurn to a tRNATrP(amber) gene and changing a tRNAG1"(UUC) gene from D. discoideum to o tRNAGl"(ochre) as well as a tRNAGl"(amber) gene.

These genes were stably integrated into the D. discoideurn genome together with a reporter gene. An actin 6::lacZ gene fusion carrying corresponding translational stop signals served as a reported.

Active P-galactosidase is expressed only in D. discoideum strains that contain, in addition to the reporter, a functional suppressor tRNA. Both amber suppressors are active in D. discoideurn without interfering significantly with cell growth and development. We failed, however, to establish cell lines containing a functional tRNAG"'(ochre) suppressor.

This may be due to the fact that nearly every message from D. discoideurn known s o far terminates with UAA. Therefore a tRNA capable of reading this termination codon may not be compatible with cell growth.

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