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Nucleic Acids Research, 1985, Vol. 13, No. 2 435-447
© 1985

Articles

Point mutations in the 5' ICR and anticodon region of a Drosophila tRNAArg gene decrease in vitro transcription

Thomas S. Stewart1,*, Dieter Soil1 and Stephen Sharp2

1Department of Molecular Biophysics and Biochemistry, Yale University New Haven, CT 06511 2Department of Microbiology and Molecular Genetics, California College of Medicine, University of California Irvine, CA 92717, USA

Received October 1, 1984. Revised December 18, 1984. Accepted December 18, 1984.

We have examined the effects of various nucleotide substitutions in a Drosophila tRNAArg gene on in vitro transcription and stable transcription complex formation in Drosophila KcO and HeLa cell extracts. Substitutions in positions encoding the invariant G18 and G19 residues resulted in decreased transcription, however, the moderate decreases indicate that these nucleotides are not obligatory promoter recognition sites. An A21 to C21 mutation had no effect on transcription levels using homologous extract however, this mutant displayed decreased transcriptional abilities in HeLa cell extract. Nucleotide substitutions within the sequence encoding the anticodon led to a decrease in the transcription activity but not in the ability to form a stable transcription complex.

*Present address: School of Biochemistry, University of New South Wales, P.O. Box 1, Kensington, 2033 N.S.W., Australia


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