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Specificity of the photo reaction of 4' hydroxymethyl 4 5' 8 trimethyl psoralen with rna identification of reactive sites in escherichia coli phenyl alanine accepting transfer rna


, : Specificity of the photo reaction of 4' hydroxymethyl 4 5' 8 trimethyl psoralen with rna identification of reactive sites in escherichia coli phenyl alanine accepting transfer rna. Biochemistry 21(6): 1357-1363

To test the potential of psoralen photoaddition for the probing of RNA conformation at sequence resolution, the specificity of the reaction of 4'-(hydroxymethyl)-4,5',8-trimethylpsoralen (HMT) with E. coli tRNAPhe was analyzed. The sites of HMT covalent addition were identified by a combination of analytical techniques involving chemical cleavage of the tRNAPhe molecule at the m7G [7-methylguanine] site and gel electrophoresis of RNase T1 digests together with paper electrophoretic characterization of HMT-modified nucleotides and oligonucleotides. Advantage was taken of the alteration of the cleavage rate of pancreatic RNase adjacent to a photoadduct. HMT photoaddition showed a very high preference for uracil residues. Important differences in HMT photoreactivity were observed for various U sites of the tRNAPhe molecule. Reactivity of specific bases was correlated with partial melting of the molecule. There appears to be a strong preference of the photoreactive probe for a loose helical conformation as compared with a tight helix, whereas a random coil appears poorly reactive.

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Related references

Bachellerie J P.; Hearst J.E., 1980: Identification of hydroxymethyl trimethyl psoralen binding sites in escherichia coli phenyl alanyl transfer rna. Federation Proceedings 39(6): ABSTRACT 2194

Bachellerie, J.P.; Hearst, J.E., 1982: Specificity of the photoreaction of 4'-(hydroxymethyl)-4,5',8-trimethylpsoralen with ribonucleic acid. Identification of reactive sites in Escherichia coli phenylalanine-accepting transfer ribonucleic acid. In order to test the potential of psoralen photoaddition for the probing of RNA conformation at sequence resolution, we have analyzed the specificity of the reaction of 4'-(hydroxymethyl)-4,5',8-trimethylpsoralen (HMT) with Escherichia c...

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Prudchenko A.T., 1971: Investigation of the specificity of phenyl alanyl transfer rna synthetase from escherichia coli mre 600 with respect to fluorinated analogues of phenyl alanine. Doklady Akademii Nauk Sssr Biochemistry Section-6: 348-350,

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