EF-Tu dynamics during pre-translocation complex formation: EF-Tu•GDP exits the ribosome via two different pathways

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EF-Tu dynamics during pre-translocation complex formation : EF-Tu•GDP exits the ribosome via two different pathways. / Liu, Wei; Chen, Chunlai; Kavaliauskas, Darius; Knudsen, Charlotte R.; Goldman, Yale E.; Cooperman, Barry S.

I: Nucleic Acids Research, Bind 43, Nr. 19, 2015, s. 9519-9528.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Harvard

Liu, W, Chen, C, Kavaliauskas, D, Knudsen, CR, Goldman, YE & Cooperman, BS 2015, 'EF-Tu dynamics during pre-translocation complex formation: EF-Tu•GDP exits the ribosome via two different pathways', Nucleic Acids Research, bind 43, nr. 19, s. 9519-9528. https://doi.org/10.1093/nar/gkv856

APA

Liu, W., Chen, C., Kavaliauskas, D., Knudsen, C. R., Goldman, Y. E., & Cooperman, B. S. (2015). EF-Tu dynamics during pre-translocation complex formation: EF-Tu•GDP exits the ribosome via two different pathways. Nucleic Acids Research, 43(19), 9519-9528. https://doi.org/10.1093/nar/gkv856

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Author

Liu, Wei ; Chen, Chunlai ; Kavaliauskas, Darius ; Knudsen, Charlotte R. ; Goldman, Yale E. ; Cooperman, Barry S. / EF-Tu dynamics during pre-translocation complex formation : EF-Tu•GDP exits the ribosome via two different pathways. I: Nucleic Acids Research. 2015 ; Bind 43, Nr. 19. s. 9519-9528.

Bibtex

@article{ce50266f2e184487bef788df0a9868b4,
title = "EF-Tu dynamics during pre-translocation complex formation: EF-Tu•GDP exits the ribosome via two different pathways",
abstract = "The G-protein EF-Tu, which undergoes a major conformational change when EF-Tu center dot GTP is converted to EF-Tu center dot GTP, forms part of an aminoacyl(aa)-tRNA center dot EF-Tu center dot GTP ternary complex (TC) that accelerates the binding of aa-tRNA to the ribosome during peptide elongation. Such binding, placing a portion of EF-Tu in contact with the GTPase Associated Center (GAC), is followed by GTP hydrolysis and Pi release, and results in formation of a pretranslocation (PRE) complex. Although tRNA movement through the ribosome during PRE complex formation has been extensively studied, comparatively little is known about the dynamics of EF-Tu interaction with either the ribosome or aa-tRNA. Here we examine these dynamics, utilizing ensemble and single molecule assays employing fluorescent labeled derivatives of EF-Tu, tRNA, and the ribosome to measure changes in either FRET efficiency or fluorescence intensity during PRE complex formation. Our results indicate that ribosome-bound EF-Tu separates from the GAC prior to its full separation from aa-tRNA, and suggest that EF-Tu center dot GDP dissociates from the ribosome by two different pathways. These pathways correspond to either reversible EF-Tu center dot GTP dissociation from the ribosome prior to the major conformational change in EF-Tu that follows GTP hydrolysis, or irreversible dissociation after or concomitant with this conformational change.",
keywords = "AMINOACYL-TRANSFER-RNA, ELONGATION-FACTOR TU, CRYSTAL-STRUCTURE, ESCHERICHIA-COLI, GTP HYDROLYSIS, INORGANIC-PHOSPHATE, BACTERIAL RIBOSOME, INITIATION COMPLEX, PROTEIN-SYNTHESIS, STRUCTURAL BASIS",
author = "Wei Liu and Chunlai Chen and Darius Kavaliauskas and Knudsen, {Charlotte R.} and Goldman, {Yale E.} and Cooperman, {Barry S.}",
year = "2015",
doi = "10.1093/nar/gkv856",
language = "English",
volume = "43",
pages = "9519--9528",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "19",

}

RIS

TY - JOUR

T1 - EF-Tu dynamics during pre-translocation complex formation

T2 - EF-Tu•GDP exits the ribosome via two different pathways

AU - Liu, Wei

AU - Chen, Chunlai

AU - Kavaliauskas, Darius

AU - Knudsen, Charlotte R.

AU - Goldman, Yale E.

AU - Cooperman, Barry S.

PY - 2015

Y1 - 2015

N2 - The G-protein EF-Tu, which undergoes a major conformational change when EF-Tu center dot GTP is converted to EF-Tu center dot GTP, forms part of an aminoacyl(aa)-tRNA center dot EF-Tu center dot GTP ternary complex (TC) that accelerates the binding of aa-tRNA to the ribosome during peptide elongation. Such binding, placing a portion of EF-Tu in contact with the GTPase Associated Center (GAC), is followed by GTP hydrolysis and Pi release, and results in formation of a pretranslocation (PRE) complex. Although tRNA movement through the ribosome during PRE complex formation has been extensively studied, comparatively little is known about the dynamics of EF-Tu interaction with either the ribosome or aa-tRNA. Here we examine these dynamics, utilizing ensemble and single molecule assays employing fluorescent labeled derivatives of EF-Tu, tRNA, and the ribosome to measure changes in either FRET efficiency or fluorescence intensity during PRE complex formation. Our results indicate that ribosome-bound EF-Tu separates from the GAC prior to its full separation from aa-tRNA, and suggest that EF-Tu center dot GDP dissociates from the ribosome by two different pathways. These pathways correspond to either reversible EF-Tu center dot GTP dissociation from the ribosome prior to the major conformational change in EF-Tu that follows GTP hydrolysis, or irreversible dissociation after or concomitant with this conformational change.

AB - The G-protein EF-Tu, which undergoes a major conformational change when EF-Tu center dot GTP is converted to EF-Tu center dot GTP, forms part of an aminoacyl(aa)-tRNA center dot EF-Tu center dot GTP ternary complex (TC) that accelerates the binding of aa-tRNA to the ribosome during peptide elongation. Such binding, placing a portion of EF-Tu in contact with the GTPase Associated Center (GAC), is followed by GTP hydrolysis and Pi release, and results in formation of a pretranslocation (PRE) complex. Although tRNA movement through the ribosome during PRE complex formation has been extensively studied, comparatively little is known about the dynamics of EF-Tu interaction with either the ribosome or aa-tRNA. Here we examine these dynamics, utilizing ensemble and single molecule assays employing fluorescent labeled derivatives of EF-Tu, tRNA, and the ribosome to measure changes in either FRET efficiency or fluorescence intensity during PRE complex formation. Our results indicate that ribosome-bound EF-Tu separates from the GAC prior to its full separation from aa-tRNA, and suggest that EF-Tu center dot GDP dissociates from the ribosome by two different pathways. These pathways correspond to either reversible EF-Tu center dot GTP dissociation from the ribosome prior to the major conformational change in EF-Tu that follows GTP hydrolysis, or irreversible dissociation after or concomitant with this conformational change.

KW - AMINOACYL-TRANSFER-RNA

KW - ELONGATION-FACTOR TU

KW - CRYSTAL-STRUCTURE

KW - ESCHERICHIA-COLI

KW - GTP HYDROLYSIS

KW - INORGANIC-PHOSPHATE

KW - BACTERIAL RIBOSOME

KW - INITIATION COMPLEX

KW - PROTEIN-SYNTHESIS

KW - STRUCTURAL BASIS

U2 - 10.1093/nar/gkv856

DO - 10.1093/nar/gkv856

M3 - Journal article

C2 - 26338772

VL - 43

SP - 9519

EP - 9528

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 19

ER -