STUDIES ON THE CHARACTERISTIC MESSENGER RNA IN THE SYSTEM INDUCING CATECHOL OXIDIZING ENZYMES IN PSEUDOMONAS EFFUSA.
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An aryl trifluoromethyl diazirine photoreactive derivative was attached to the 2-thiocytidine residue at position 32 of tRNA(IArg) and this derivatized tRNA was bound to Escherichia coli 70S ribosomes. After irradiation at 350 nm the site of cross-linking to the 16S RNA was analyzed by our standard procedures and found to lie within the secondary structural element comprising bases 956-983; this region contains two modified nucleotides at positions 966 and 967. Similarly, an aryl azido photoreactive derivative was attached to the phenylalanine residue of Phe-tRNA(Phe), and the derivatized aminoacyl tRNA was bound to the ribosome either at the A- or the P-site. In both cases, after irradiation at 250 nm, the cross-link site was localized to position 2439 of the 23S RNA; in the secondary structure of the latter the neighboring nucleotide 2442 is base-paired to a modified nucleotide at position 2069. Taken together with other cross-linking data, these results now directly implicate a total of 27 out of the 29 modified nucleotides in E. coli 16S and 23S RNA as lying within or close to the functional center of the ribosome.
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A transfer RNA molecule accepts a particular amino acid from an activating enzyme and subsequently transfers it to the end of a growing peptide chain, in association with ribosomes and messenger RNA. The acceptor and transfer functions of transfer RNA differ in their sensitivity to damage by bromination, and thus appear to comprise different areas on the transfer RNA molecule.
A brief exposure of Escherichia coli cells to an indutcer for beta- galactosidase results in the production of a messenger RNA which subsequently expresses itself as beta-galactosidase. Ultraviolet irradiation of cells after exposure to the inducer results in a decrease in the amount of beta-galactosidase formed. The messenger RNA formed during the brief exposure to inducer is evidently inactivated by the ultraviolet radiation. The decay of messenger RNA activity in irradiated cells has the same kinetics as that observed in unirradiated cells.
The complete nucleotide sequence of an alanine transfer RNA, isolated from yeast, has been determined. This is the first nucleic acid for which the structure is known.
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