Nucleic Acids Research, 1984, Vol. 12, No. 8 3521-3534
© 1984
MOLECULAR BIOLOGY |
Novel E. coli mutants deficient in blosynthesis of 5-methylamlnomethyl-2-thiouridine
Department of Microbiology, New York Medical College Valhalla, NY 10595, USA
Received December 15, 1983. Revised April 5, 1984. Accepted April 27, 1984.
Novel E. coli mutants deficient in biosynthesis of 5-methylaminomethyl-2- thiouridine were isolated based on a phenotype of reduced readthrough at UAG codons. They define 2 new loci trmE and trmF, near 83' on the E. coli map. These mutants are different from strains carrying trmC mutations, which are known to confer a methylation deficiency in biosynthesis of 5-methylaminomethyl-2-thiouridine. tRNA from mutants carrying trmE or trmF mutations was shown to carry 2-thiouridine instead of 5-methylaminomethyl-2-thiouridine. This deficiency affects the triplet binding properties of the mutant tRNA. Our results suggest that the 5-methylaminomethyl group stabilizes the basepairing this modified nucleotide with G, most likely through direct interaction with the ribosomal binding site(s).
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