Probing the structure of the Escherichia coli 10Sa RNA (tmRNA)

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Publication Type Journal Article
School or College School of Medicine; College of Engineering; College of Science
Department Human Genetics; Howard Hughes Medical Institute; Bioengineering; Biology
Creator Gesteland, Raymond F.; Atkins, John F.; Felden, Brice; McCutcheon, John P.
Other Author Himeno, Hyouta; Muto, Akira
Title Probing the structure of the Escherichia coli 10Sa RNA (tmRNA)
Date 1997
Description The conformation of the Escherichia coli 10Sa RNA (tmRNA) in solution was investigated using chemical and enzymatic probes. Single- and double-stranded domains were identified by hydrolysis of tmRNA in imidazole buffer and by lead(II)-induced cleavages. Ribonucleases T1 and S1 were used to map unpaired nucleotides and ribonuclease V1 was used to identify paired bases or stacked nucleotides. Specific atomic positions of bases were probed with dimethylsulfate, a carbodiimide, and diethylpyrocarbonate. Covariations, identified by sequence alignment with nine other tmRNA sequences, suggest the presence of several tertiary interactions, including pseudoknots. Temperature-gradient gel electrophoresis experiments showed structural transitions of tmRNA starting around 40 degrees C, and enzymatic probing performed at selected temperatures revealed the progressive melting of several predicted interactions. Based on these data, a secondary structure is proposed, containing two stems, four stem-loops, four pseudoknots, and an unstable structural domain, some connected by single-stranded A-rich sequence stretches. A tRNA-like domain, including an already reported acceptor branch, is supported by the probing data. A second structural domain encompasses the coding sequence, which extends from the top of one stem-loop to the top of another, with a 7-nt single-stranded stretch between. A third structural module containing pseudoknots connects and probably orients the tRNA-like domain and the coding sequence. Several discrepancies between the probing data and the phylogeny suggest that E. coli tmRNA undergoes a conformational change.
Type Text
Publisher Cold Spring Harbor Lab Press
First Page 89
Last Page 103
Subject Covariation; Pseudoknot; Structural Probing; 10Sa RNA; tmRNA
Subject MESH Base Sequence; Escherichia coli; Molecular Sequence Data; RNA, Transfer
Language eng
Bibliographic Citation Felden, B., Himeno, H., Muto, A., McCutcheon, J.P., Atkins, J.F., and Gesteland, R.F. 1997. Probing the structure of the Escherichia coli 10Sa RNA (tmRNA). RNA 3:89-103
Rights Management © Cold Spring Harbor Laboratory Press-CSHL Press
Format Medium application/pdf
Identifier ir-main,12751
ARK ark:/87278/s6pk10cw
Setname ir_uspace
ID 703796
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