Effect of ultraviolet irradiation on 30-S ribosomal subunits. Identification of the RNA region crosslinked to protein S7

Eur J Biochem. 1980 Feb;104(1):255-62. doi: 10.1111/j.1432-1033.1980.tb04423.x.

Abstract

The effects of ultraviolet irradiation on Escherichia coli 30-S ribosomal subunits were studied. At the doses of radiation used in this work (0-4.5 x 10(5) quanta/30-S subunit), only protein S7 was found to be significantly crosslinked to the 16-S RNA. In conditions where 25% of the protein was covalently crosslinked, the ability of the irradiated 30-S subunits to reassociate with 50-S subunits and their activity in polyphenylalanine synthesis decreased strongly. Similar results were obtained by irradiation with a germicide lamp (254 nm) or with a monochromatic ultraviolet light at 248 nm. No additional proteins were crosslinked to the 16-S RNA by irradiating 30-S subunits depleted in protein S1 or 70-S ribosomes. The covalent complex of 16-S RNA and protein S7 was isolated and digested by T1 ribonuclease. The oligonucleotide remaining attached to the crosslinked protein was characterised as A-C-C-U-C-G [position 1261 - 1266, see the sequence published by Carbon et al. (1979) Eur. J. Biochem. 160, 399-410]. Analysis of this fragment suggests that protein S7 was linked to the cytosine at position 1265 in the RNA sequence.

MeSH terms

  • Dose-Response Relationship, Radiation
  • Escherichia coli / radiation effects*
  • Macromolecular Substances
  • Oligoribonucleotides
  • Protein Binding
  • RNA, Ribosomal / radiation effects*
  • Ribosomal Proteins / radiation effects*
  • Ribosomes / radiation effects*
  • Ultraviolet Rays*

Substances

  • Macromolecular Substances
  • Oligoribonucleotides
  • RNA, Ribosomal
  • Ribosomal Proteins