The binding of the pyrophosphoryl transferase and the elongation factor Tu and G to ribosomes from <i>Escherichia coli</i>

Kleinert, Ursula; Richter, Dietmar

doi:10.1016/0014-5793(75)80989-6

Cited by 20 publications

(9 citation statements)

References 16 publications

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“…3H-Labeled stringent factor was prepared by the reductive methylation method (ref. 9; specific activity 0.5-5.0 Ci/mol).…”

mentioning

confidence: 99%

Stringent factor from Escherichia coli directs ribosomal binding and release of uncharged tRNA.

Richter¹

1976

Proc. Natl. Acad. Sci. U.S.A.

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Uncharged tRNA is preferentially bound to the peptidyl site of the ribosome in the absence of stringent factor, but in its presence is directed to the acceptor site. The synthesis of pppGpp and ppGpp is initiated by tRNA bound in the acceptor but not in the peptidyl site. In this reaction, tRNA is not permanently attached to the acceptor site. Uncharged [32PltRNA but not 3H-labeled stringent factor is released from the ribosome after each round of stringent factor-dependent hydrolysis of ATP. ATP-32PPi-exchange experiments reveal that exchange is independent of the presence of GTP but strongly enhanced by the addition of stringent factor and tRNA. The tRNA release is suppressed when ATP is replaced by #,'y-imido adenosine 5'-triphosphate, 5'-AMP, or GTP.

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“…3H-Labeled stringent factor was prepared by the reductive methylation method (ref. 9; specific activity 0.5-5.0 Ci/mol).…”

mentioning

confidence: 99%

Stringent factor from Escherichia coli directs ribosomal binding and release of uncharged tRNA.

Richter¹

1976

Proc. Natl. Acad. Sci. U.S.A.

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“…The preparation of Escherichia coli elongation factors followed instructions given by Arai et al [3] . Labeling of EF-Tu by reductive alkylation was performed as described by Kleinert and Richter [4]. E. coli ribosomes were prepared and washed fourtimes with 0.5 M NH4Cl as described previously [S] .…”

Section: Methodsmentioning

confidence: 99%

Interchangeability of elongation factor‐Tu and elongation factor‐1 in aminoacyl‐tRNA binding to 70 S and 80 S ribosomes

1977

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“…The purity of the preparation was not less than 98':/, according to electrophoretic data in sodium-dodecylsulfate/polyacrylamide gel and N-terminal analysis. The 3H-labelled EF-G was obtained by a reductive methylation with NaB3H4 according to Kleinert and Richter [7].…”

Section: Methodsmentioning

confidence: 99%

Localization of the GTP‐Binding Site in the Ribosome · Elongation‐Factor‐G · GTP Complex

Girshovich¹,

Pozdnyakov²,

Ovchinnikov³

1976

European Journal of Biochemistry

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For localization of the GTP‐binding center in the Escherichia coli MRE‐600 ribosome elongation‐factor‐G GTP system a method of photo‐affinity labelling is used with two types of radioactive analogs oI GTP with a photo‐activated group attached to ribose or to γ‐phosphate residues of a nucleotide molecule: (2‐nitro,4‐azidobenzoyl)hydrazone of periodate‐oxidized GTP or guanosine 5′‐(α,β‐methylene)triphosphate and the γ‐(4‐azidobenzyl)amide of GTP. These photo‐analogs form a specific ternary complex with the ribosome and elongation factor G (EF‐G). Separately neither the ribosome tor the EF‐G bind analogs. The formation of the ternary complex with analogs is inhibited by an excess amount of the native nucleotide. The ribose analog of GTP is hydrolyzed by the EF‐G‐dependent GTPase, but the γ‐amide of GTP is not. Irradiation of complexes containing photo‐analogs leads to a highly specific labelling of EF‐G, while ribosome labelling is insignificant and non‐specific. The presence of an excess amount of the native nucleotide inhibits photo‐labelling of EF‐G without significant changes in the labelling of ribosomes. The labelling of free EF‐G is low. Thus, within the ternary complex the GTP‐binding center is localized on the EF‐G. Fusidic acid does not influence the binding of GTP γ‐amide in a ternary complex, but blocks the specific labelling of the EF‐G with no effect on the non‐specific labelling of ribosomes. A hypothesis is proposed on the direct contact of this antibiotic with a GTP molecule in the GTP‐binding center of EF‐G.

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The binding of the pyrophosphoryl transferase and the elongation factor Tu and G to ribosomes from Escherichia coli

Cited by 20 publications

References 16 publications

Stringent factor from Escherichia coli directs ribosomal binding and release of uncharged tRNA.

Stringent factor from Escherichia coli directs ribosomal binding and release of uncharged tRNA.

Interchangeability of elongation factor‐Tu and elongation factor‐1 in aminoacyl‐tRNA binding to 70 S and 80 S ribosomes

Localization of the GTP‐Binding Site in the Ribosome · Elongation‐Factor‐G · GTP Complex

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