Ribosomal proteins at the stalk region modulate functional rRNA structures in the GTPase center.
Replacement of the L10.L7/L12 protein complex and L11 in Escherichia coli ribosomes with the respective rat counterparts P0.P1/P2 and eukaryotic L12 causes conversion of ribosomal specificity for elongation factors from prokaryotic elongation factor (EF)-Tu/EF-G to eukaryotic EF (eEF)-1alpha/eEF-2. Here we have investigated the effects of protein replacement on the structure and function of two rRNA domains around positions 1070 and 2660 (sarcin/ricin loop) of 23 S rRNA. Protein replacement at the 1070 region in E. coli 50 S subunits was demonstrated by chemical probing analysis. Binding of rat proteins to the 1070 region caused increased accessibility of the 2660 and 1070 regions to ligands for eukaryotic ribosomes: the ribotoxin pepocin for the 2660 region (E. coli numbering), anti-28 S autoantibody for the 1070 region, and eEF-2 for both regions. Moreover, binding of the E. coli L10.L7/L12 complex and L11 to the 1070 region was shown to be responsible for E. coli ribosomal accessibility to another ribotoxin, gypsophilin. Ribosomal proteins at the 1070 region appear to modulate the structures and functions of the 2660 and 1070 RNA regions in slightly different modes in prokaryotes and eukaryotes.
Institute of High Polymer Research, Faculty of Textile Science and Technology, Shinshu University, Ueda 386-8567, Japan. uchiumi@giptc.shinshu-u.ac.jp
Toshio
T
Uchiumi
Sachiko
S
Honma
Yaeta
Y
Endo
Akira
A
Hachimori
D000818:Animals; D004722:Endoribonucleases / Q000737:chemistry; D005656:Fungal Proteins; D020558:GTP Phosphohydrolases / Q000737:chemistry*; D009690:Nucleic Acid Conformation; D020652:Peptide Elongation Factor 2 / Q000378:metabolism; D012335:RNA, Ribosomal / Q000737:chemistry*; D051381:Rats; D012269:Ribosomal Proteins / Q000737:chemistry*; D012270:Ribosomes / Q000378:metabolism; D012276:Ricin / Q000737:chemistry
D005656:Fungal Proteins; D020652:Peptide Elongation Factor 2; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; C014801:alpha-sarcin; D012276:Ricin; D004722:Endoribonucleases; D020558:GTP Phosphohydrolases