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Chapter 29 : The States, Conformational Dynamics, and Fusidic Acid-Resistant Mutants of Elongation Factor G
Authors:
Anders Liljas1,
Ole Kristensen1,
Martin Laurberg1,
Salam Al-Karadaghi1,
Anatoly Gudkov2,
Kirill Martemyanov2,
Diarmaid Hughes3,
Ivan Nagaev3
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Affiliations:
1: Molecular Biophysics, Lund University, Box 124, SE-22100 Lund, Sweden;
2: Institute of Protein Research, Russian Academy of Sciences, Pushchino,Moscow Region, Russia;
3: Department of Cell and Molecular Biology, BMC, Uppsala University, Box 596, SE-75124 Uppsala, Sweden
Content Type: Monograph
Publication Year:
2000
Category: Microbial Genetics and Molecular Biology
Book DOI:
10.1128/9781555818142
Chapter DOI:
10.1128/9781555818142.ch29
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Abstract:
This chapter summarizes the current knowledge and proposes a structural model for the function of elongation factor G (EF-G). The large number of mutations associated with fusidic acid resistance are an essential ingredient in this analysis. Recent investigations of a mutant EF-G with a different crystal packing have led to a complete interpretation of domain III at relatively low resolution. The functional cycle of EF-G can be described as a number of states both on and off the ribosome. The different states of EF-G may not necessarily be associated with different conformations of EF-G, but to the extent that there are different conformations, they will be related to different states. The density of EF-G could be identified with difference methods and compared to the crystallographic GDP conformation. The mutant G16V is fusidic acid sensitive compared to wt Thermus thermophilus EF-G, which is relatively resistant to the antibiotic. During one of the subsequent steps, EF-G adopts an open conformation like the one observed by cryo-EM. Since it overlaps the A-site tRNA, translocation must already have occurred, as is well known from studies of fusidic acid inhibition of protein synthesis. When EF-G has dissociated, it has the intermediate GDP conformation.
Citation:
Liljas A, Kristensen O, Laurberg M, Al-Karadaghi S, Gudkov A, Martemyanov K, Hughes D, Nagaev I.
2000.
The States, Conformational Dynamics, and Fusidic Acid-Resistant Mutants of Elongation Factor G,
p 359-365.
In
Garett R, Douthwaite S, Liljas A, Matheson A, Moore P, Noller H (ed),
The Ribosome.
ASM Press, Washington, DC.
doi: 10.1128/9781555818142.ch29
Figures
The States, Conformational Dynamics, and Fusidic Acid-Resistant Mutants of Elongation Factor G
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Citation:
Liljas A, Kristensen O, Laurberg M, Al-Karadaghi S, Gudkov A, Martemyanov K, Hughes D, Nagaev I.
2000.
The States, Conformational Dynamics, and Fusidic Acid-Resistant Mutants of Elongation Factor G,
p 359-365.
In
Garett R, Douthwaite S, Liljas A, Matheson A, Moore P, Noller H (ed),
The Ribosome.
ASM Press, Washington, DC.
doi: 10.1128/9781555818142.ch29
/content/10.1128/9781555818142.chap29.fig29-1
Figure 1
Representation of some of the possible functional states of EF-G according to the classical model (a) and the model of Rodnina et al. (1997) (b). The white boxes represent the ribosome, the gray symbols represent EF-G in different states, and the black triangles represent GTP (base down) or GDP (base up).
10.1128/9781555818142/fig29-1_thmb.gif
10.1128/9781555818142/fig29-1.gif
Figure 1
Representation of some of the possible functional states of EF-G according to the classical model (a) and the model of Rodnina et al. (1997) (b). The white boxes represent the ribosome, the gray symbols represent EF-G in different states, and the black triangles represent GTP (base down) or GDP (base up).
Citation:
Liljas A, Kristensen O, Laurberg M, Al-Karadaghi S, Gudkov A, Martemyanov K, Hughes D, Nagaev I.
2000.
The States, Conformational Dynamics, and Fusidic Acid-Resistant Mutants of Elongation Factor G,
p 359-365.
In
Garett R, Douthwaite S, Liljas A, Matheson A, Moore P, Noller H (ed),
The Ribosome.
ASM Press, Washington, DC.
doi: 10.1128/9781555818142.ch29
/content/10.1128/9781555818142.chap29.fig29-2
Figure 2
Comparison of the two main conformations of EF-G observed crystallographically. The broad ribbon represents domains G, G′, and II, which here are fixed. The narrow tube represents domains III to V. The GDP conformation is shown in orange, and the new bent conformation of mutant H573A is shown in green. The red bar indicates the axis around which the rotation of about 10° occurs.
10.1128/9781555818142/fig29-2_thmb.gif
10.1128/9781555818142/fig29-2.gif
Figure 2
Comparison of the two main conformations of EF-G observed crystallographically. The broad ribbon represents domains G, G′, and II, which here are fixed. The narrow tube represents domains III to V. The GDP conformation is shown in orange, and the new bent conformation of mutant H573A is shown in green. The red bar indicates the axis around which the rotation of about 10° occurs.
Citation:
Liljas A, Kristensen O, Laurberg M, Al-Karadaghi S, Gudkov A, Martemyanov K, Hughes D, Nagaev I.
2000.
The States, Conformational Dynamics, and Fusidic Acid-Resistant Mutants of Elongation Factor G,
p 359-365.
In
Garett R, Douthwaite S, Liljas A, Matheson A, Moore P, Noller H (ed),
The Ribosome.
ASM Press, Washington, DC.
doi: 10.1128/9781555818142.ch29
/content/10.1128/9781555818142.chap29.fig29-3
Figure 3
Comparison of the structures of domains III and V. The same fold is observed. This fold, the RNA recognition motif (RRM), has also been observed in many ribosomal proteins.
10.1128/9781555818142/fig29-3_thmb.gif
10.1128/9781555818142/fig29-3.gif
Figure 3
Comparison of the structures of domains III and V. The same fold is observed. This fold, the RNA recognition motif (RRM), has also been observed in many ribosomal proteins.
Citation:
Liljas A, Kristensen O, Laurberg M, Al-Karadaghi S, Gudkov A, Martemyanov K, Hughes D, Nagaev I.
2000.
The States, Conformational Dynamics, and Fusidic Acid-Resistant Mutants of Elongation Factor G,
p 359-365.
In
Garett R, Douthwaite S, Liljas A, Matheson A, Moore P, Noller H (ed),
The Ribosome.
ASM Press, Washington, DC.
doi: 10.1128/9781555818142.ch29
/content/10.1128/9781555818142.chap29.fig29-4
Figure 4
Locations of fusidic acid-resistant mutations in EF-G (indicated in pink). The six most resistant mutants are shown in red. The right panel is an enlargement of the central region of the left panel.
10.1128/9781555818142/fig29-4_thmb.gif
10.1128/9781555818142/fig29-4.gif
Figure 4
Locations of fusidic acid-resistant mutations in EF-G (indicated in pink). The six most resistant mutants are shown in red. The right panel is an enlargement of the central region of the left panel.
Citation:
Liljas A, Kristensen O, Laurberg M, Al-Karadaghi S, Gudkov A, Martemyanov K, Hughes D, Nagaev I.
2000.
The States, Conformational Dynamics, and Fusidic Acid-Resistant Mutants of Elongation Factor G,
p 359-365.
In
Garett R, Douthwaite S, Liljas A, Matheson A, Moore P, Noller H (ed),
The Ribosome.
ASM Press, Washington, DC.
doi: 10.1128/9781555818142.ch29
/content/10.1128/9781555818142.chap29.fig29-5
Figure 5
Superposition of EF-G (van der Waals surface) in complex with GDP and the ternary complex of EF-Tu with tRNA and GDPNP (atomic model). The ternary complex has been found to have a somewhat more open conformation.
10.1128/9781555818142/fig29-5_thmb.gif
10.1128/9781555818142/fig29-5.gif
Figure 5
Superposition of EF-G (van der Waals surface) in complex with GDP and the ternary complex of EF-Tu with tRNA and GDPNP (atomic model). The ternary complex has been found to have a somewhat more open conformation.
Citation:
Liljas A, Kristensen O, Laurberg M, Al-Karadaghi S, Gudkov A, Martemyanov K, Hughes D, Nagaev I.
2000.
The States, Conformational Dynamics, and Fusidic Acid-Resistant Mutants of Elongation Factor G,
p 359-365.
In
Garett R, Douthwaite S, Liljas A, Matheson A, Moore P, Noller H (ed),
The Ribosome.
ASM Press, Washington, DC.
doi: 10.1128/9781555818142.ch29
/content/10.1128/9781555818142.chap29.fig29-6
Figure 6
Comparison of the observed conformations of EF-G with those of the ternary complex of EF-Tu. Fus, fusidic acid; kirr, kirromycin; aa-tRNA, aminoacyl-tRNA.
10.1128/9781555818142/fig29-6_thmb.gif
10.1128/9781555818142/fig29-6.gif
Figure 6
Comparison of the observed conformations of EF-G with those of the ternary complex of EF-Tu. Fus, fusidic acid; kirr, kirromycin; aa-tRNA, aminoacyl-tRNA.
Citation:
Liljas A, Kristensen O, Laurberg M, Al-Karadaghi S, Gudkov A, Martemyanov K, Hughes D, Nagaev I.
2000.
The States, Conformational Dynamics, and Fusidic Acid-Resistant Mutants of Elongation Factor G,
p 359-365.
In
Garett R, Douthwaite S, Liljas A, Matheson A, Moore P, Noller H (ed),
The Ribosome.
ASM Press, Washington, DC.
doi: 10.1128/9781555818142.ch29
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