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Nucleic Acids Research, 1984, Vol. 12, No. 3 1473-1487
© 1984

MOLECULAR BIOLOGY

Binding of tRNA alters the chemical accessibility of nucleotides within the large ribosomal RNAs of E. coli ribosomes

N. Meier and R. Wagner

Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann Ihnestrasse 63-73, 1000 Berlin-Dahlem, FRG

Received October 20, 1983. Revised December 15, 1983. Accepted December 15, 1983.

Functionally active 70S ribosomes were chemically modified with dimethylsulfate (DMS) in the presence and absence of bound tRNA. The ribosomal 16S RNA and 23S RNA were extracted, separat ed and labeled radioactively at their 3'-ends. DMS modification sites within the last 200 nucleotides from the 3'-ends were investigated on sequencing gels, after borohydride reduction and aniline catalyzed strand scission of the isolated RNA's.

tRNA binding caused enhanced reactivity at 9 nucleotide positions while three sites showed decreased reactivity in the 16S RNA. The effects of bound tRNA on the modification of 23S RNA were limited. Only one enhancement was observed in the pre sence of bound tRNA. mRNA binding alone showed two more sites with enhanced reactivity, however. The results are consistent with the view that the sequence 1400–1500 of the 16S RNA plays an important functional role in the translating ribosome and possibly constitutes part of the tRNA binding site.


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