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The Binding Sites of Class I Release Factor (eRF1) Toward Class II Release Factor (eRF3) in Euplotes octocarinatus

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Abstract

The C domain of eRF1 interacts with the C domain of eRF3, and the binding of both factors is essential for fast kinetics of the termination of protein translation. Analysis by computational simulation demonstrated that several peptides involved in Eo-eRF1/Eo-eRF3 interaction directly. Among these peptides, the two motifs GVEDT and GFGG were highly conserved, while the fragment aa338-346 of Eo-eRF1a/b was variable. In additional, I290 and D293 of Eo-eRF1 were also highly conserved. By the site-directed mutagenesis and pull-down analysis, the amino acid D293 in Eo-eRF1bC domain was conformed playing an important role in eRF1–eRF3 interaction. Eo-eRF1a and Eo-eRF1b may select different manners to interact with Eo-eRF3. These studies contribute to the better understanding the mode of eRF1–eRF3 interaction.

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Abbreviations

eRF1:

Class I polypeptide release factor in eukaryotes

eRF3:

Class II polypeptide release factor in eukaryotes

Bj-eRF1:

Class I polypeptide release factor in Blepharisma japonicum

Eo-eRF1a:

Class I polypeptide release factor a in Euplotes octocarinatus

Eo-eRF1b:

Class I polypeptide release factor b in E. octocarinatus

Hs-eRF1:

Class I polypeptide release factor in Homo sapiens

Bj-eRF1C:

C domain of Bj-eRF1 (274–436)

Eo-eRF1aC:

C domain of Eo-eRF1a (270–446)

Eo-eRF1bC:

C domain of Eo-eRF1b (270–437)

Eo-eRF3:

Class II polypeptide release factor in E. octocarinatus

Eo-eRF3Cm6:

Truncated peptide of Eo-eRF3 (640–723)

Bj-eRF1C/Eo-eRF3Cm6:

Heterodimeric complex of Bj-eRF1C and Eo-eRF3Cm6

Eo-eRF1aC/Eo-eRF3Cm6:

Heterodimeric complex of Eo-eRF1aC and Eo-eRF3Cm6

Eo-eRF1bC/Eo-eRF3Cm6:

Heterodimeric complex of Eo-eRF1bC and Eo-eRF3Cm6

MD:

Molecular dynamics

RLS:

Resonance light scattering

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Acknowledgments

This work was supported by grants from the National Natural Scientific Foundation of China (Nos. 31071294, 30770294, and 30770295).

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Authors and Affiliations

  1. Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan, 030006, China

    Jie Chen, Yue-jun Fu & Ai-hua Liang

  2. Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Molecular Science, Shanxi University, Taiyuan, 030006, China

    Bing-sheng Yang & Yan-bo Wu

Authors
  1. Jie Chen
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  2. Yue-jun Fu
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  3. Bing-sheng Yang
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  4. Yan-bo Wu
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  5. Ai-hua Liang
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Corresponding author

Correspondence to Ai-hua Liang.

Electronic Supplementary Material

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Table S1

The parameters of hydrogen bond in Eo-eRF1bC/Eo-eRF3Cm6 complex (DOCX 12 kb)

Table S2

The parameters of hydrogen bond in Eo-eRF1aC/Eo-eRF3Cm6 complex (DOCX 12 kb)

Table S3

The parameters of hydrogen bond in Bj-eRF1C/Eo-eRF3Cm6 complex (DOCX 11 kb)

Figure S1

The conformers of minidomain and the site D297 in eRF1C/Eo-eRF3Cm6 complexes (DOC 1977 kb)

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Chen, J., Fu, Yj., Yang, Bs. et al. The Binding Sites of Class I Release Factor (eRF1) Toward Class II Release Factor (eRF3) in Euplotes octocarinatus . Appl Biochem Biotechnol 165, 1507–1518 (2011). https://doi.org/10.1007/s12010-011-9371-3

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