Altered lead(II)-cleavage pattern of free Phe-tRNA(Phe) and Phe-tRNA(Phe) in ternary complex with EF-Tu:GTP

TY - JOUR

T1 - Altered lead(II)-cleavage pattern of free Phe-tRNA(Phe) and Phe-tRNA(Phe) in ternary complex with EF-Tu:GTP

AU - Otzen, D. E.

AU - Barciszewski, J.

AU - Clark, B. F.C.

PY - 1993/1/1

Y1 - 1993/1/1

N2 - Pb2+ ions in sub-millimolar concentrations are known to cleave internucleotide bonds of phenylalanine-specific transfer RNA (tRNA(Phe)) from Saccharomyces. cerevisiae specifically between nucleotides D17 and G18 in the D-loop, with additional minor cleavages after D16 and G15. This makes lead(II) a sensitive structural probe for correct folding of tRNA(Phe). In the present paper we use Pb2+ ions as a functional probe to determine whether this part of tRNA is protected by the Escherichia coli elongation factor EF-Tu in the ternary complex formed between Phe-tRNA(Phe) and EF-Tu.GTP. Our results show that for tRNA in complex with EF-Tu:GTP, the phosphodiester bond after D17 is cleaved, yet the phosphodiester bonds after D16 and G15 are not. To our knowledge, this is the first time that Pb2+ ions, bound at a specific site in tRNA, have been used both to investigate the correct folding of tRNA in complex, and to footprint a functional complex with components whose individual structures are known.

AB - Pb2+ ions in sub-millimolar concentrations are known to cleave internucleotide bonds of phenylalanine-specific transfer RNA (tRNA(Phe)) from Saccharomyces. cerevisiae specifically between nucleotides D17 and G18 in the D-loop, with additional minor cleavages after D16 and G15. This makes lead(II) a sensitive structural probe for correct folding of tRNA(Phe). In the present paper we use Pb2+ ions as a functional probe to determine whether this part of tRNA is protected by the Escherichia coli elongation factor EF-Tu in the ternary complex formed between Phe-tRNA(Phe) and EF-Tu.GTP. Our results show that for tRNA in complex with EF-Tu:GTP, the phosphodiester bond after D17 is cleaved, yet the phosphodiester bonds after D16 and G15 are not. To our knowledge, this is the first time that Pb2+ ions, bound at a specific site in tRNA, have been used both to investigate the correct folding of tRNA in complex, and to footprint a functional complex with components whose individual structures are known.

UR - http://www.scopus.com/inward/record.url?scp=0027520018&partnerID=8YFLogxK

M3 - Journal article

C2 - 8260950

AN - SCOPUS:0027520018

SN - 0158-5231

VL - 31

SP - 95

EP - 103

JO - Biochemistry and Molecular Biology International

JF - Biochemistry and Molecular Biology International

IS - 1

ER -