On the interaction of ribosomal protein L5 with 5S rRNA

Kenta Iwasaki, Shingo Kikukawa, Shunsuke Kawamura, Yoshiaki Kouzuma, Isao Tanaka, Makoto Kimura

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Ribosomal protein L5, a 5S rRNA binding protein in the large subunit, is composed of a five-stranded antiparallel β-sheet and four α-helices, and folds in a way that is topologically similar to the ribonucleprotein (RNP) domain [Nakashima et al., RNA 7, 692–701, 2001]. The crystal structure of ribosomal protein L5 (BstL5) from Bacillus stearothermophilus suggests that a concave surface formed by an anti-parallel β-sheet and long loop structures are strongly involved in 5S rRNA binding. To identify amino acid residues responsible for 5S rRNA binding, we made use of Ala-scanning mutagenesis of evolutionarily conserved amino acids occurred at β-strands and loop structures in BstL5. The mutation of Lys33 at the β1-strand caused a significant reduction in 5S rRNA binding. In addition, the Arg92, Phe122, and Glu134 mutations on the β2-strand, the α3-β4 loop, and the β4-β5 loop, respectively, resulted in a moderate decrease in the 5S rRNA binding affinity. In contrast, mutation of the conserved residue Pro65 at the β2-strand had little effect on the 5S rRNA binding activity. These results, taken together with previous results, identified Lys33, Asn37, Gln63, and Thr90 on the β-sheet structure, and Phe77 at the β2-β3 loop as critical residues for the 5S rRNA binding. The contribution of these amino acids to 5S rRNA binding was further quantitatively evaluated by surface plasmon resonance (SPR) analysis by the use of BIAcore. The results showed that the amino acids on the β-sheet structure are required to decrease the dissociation rate constant for the BstL5-5S rRNA complex, while those on the loops are to increase the association rate constant for the BstL5-5S rRNA interaction.

Original languageEnglish
Pages (from-to)103-109
Number of pages7
JournalBioscience, Biotechnology and Biochemistry
Volume66
Issue number1
DOIs
Publication statusPublished - Jan 1 2002

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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  • Cite this

    Iwasaki, K., Kikukawa, S., Kawamura, S., Kouzuma, Y., Tanaka, I., & Kimura, M. (2002). On the interaction of ribosomal protein L5 with 5S rRNA. Bioscience, Biotechnology and Biochemistry, 66(1), 103-109. https://doi.org/10.1271/bbb.66.103