The Co-chaperone Cns1 and the Recruiter Protein Hgh1 Link Hsp90 to Translation Elongation via Chaperoning Elongation Factor 2

Schopf, Florian H.; Huber, E.M.; Dodt, Christopher; Lopez, Abraham; Biebl, Maximilian M.; Rutz, Daniel; Mühlhofer, Moritz; Richter, Gesa; Madl, Tobias; Sattler, Michael; Groll, M.; Büchner, Johannes

doi:10.1016/j.molcel.2019.02.011

Cited by 26 publications

(28 citation statements)

References 92 publications

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“…EF-G is highly conserved across all kingdoms of life. Efficient folding of the eukaryotic EF-G ortholog eEF2 requires a specialized chaperone, Hgh1, that likely binds cotranslationally, stabilizes domain III, and recruits additional chaperones, including the TRiC chaperonin and possibly Hsp90, to the nascent polypeptide (61, 62). Bacteria lack Hgh1 and likely use other chaperone systems to reduce misfolding during synthesis.…”

Section: Discussionmentioning

confidence: 99%

Energetic dependencies dictate folding mechanism in a complex protein

Liu

Chen

Kaiser

2019

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

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Large proteins with multiple domains are thought to fold cotranslationally to minimize interdomain misfolding. Once folded, domains interact with each other through the formation of extensive interfaces that are important for protein stability and function. However, multidomain protein folding and the energetics of domain interactions remain poorly understood. In elongation factor G (EF-G), a highly conserved protein composed of 5 domains, the 2 N-terminal domains form a stably structured unit cotranslationally. Using single-molecule optical tweezers, we have defined the steps leading to fully folded EF-G. We find that the central domain III of EF-G is highly dynamic and does not fold upon emerging from the ribosome. Surprisingly, a large interface with the N-terminal domains does not contribute to the stability of domain III. Instead, it requires interactions with its folded C-terminal neighbors to be stably structured. Because of the directionality of protein synthesis, this energetic dependency of domain III on its C-terminal neighbors disrupts cotranslational folding and imposes a posttranslational mechanism on the folding of the C-terminal part of EF-G. As a consequence, unfolded domains accumulate during synthesis, leading to the extensive population of misfolded species that interfere with productive folding. Domain III flexibility enables large-scale conformational transitions that are part of the EF-G functional cycle during ribosome translocation. Our results suggest that energetic tuning of domain stabilities, which is likely crucial for EF-G function, complicates the folding of this large multidomain protein.

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Section: Discussionmentioning

confidence: 99%

Energetic dependencies dictate folding mechanism in a complex protein

Liu

Chen

Kaiser

2019

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

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“…In this case, the ribosome, together with trigger factor (TF), a cotranslationally acting chaperone, aids early folding steps to establish the correct path for folding [101]. Notably, in vivo folding of the eukaryotic homolog of EF-G, eEF2, requires the help of chaperones [102,103]. Interestingly, recent work on the cotranslational folding of domain III of EF-G shows that this domain is not stabilized by its N-terminal neighbors (domain G and domain II) and requires interactions with the C-terminal domains (domains IV and V) to adopt a stable structure [104].…”

Section: Multidomain Protein Foldingmentioning

confidence: 99%

Cotranslational Folding of Proteins on the Ribosome

2020

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Many proteins in the cell fold cotranslationally within the restricted space of the polypeptide exit tunnel or at the surface of the ribosome. A growing body of evidence suggests that the ribosome can alter the folding trajectory in many different ways. In this review, we summarize the recent examples of how translation affects folding of single-domain, multiple-domain and oligomeric proteins. The vectorial nature of translation, the spatial constraints of the exit tunnel, and the electrostatic properties of the ribosome-nascent peptide complex define the onset of early folding events. The ribosome can facilitate protein compaction, induce the formation of intermediates that are not observed in solution, or delay the onset of folding. Examples of single-domain proteins suggest that early compaction events can define the folding pathway for some types of domain structures. Folding of multi-domain proteins proceeds in a domain-wise fashion, with each domain having its role in stabilizing or destabilizing neighboring domains. Finally, the assembly of protein complexes can also begin cotranslationally. In all these cases, the ribosome helps the nascent protein to attain a native fold and avoid the kinetic traps of misfolding.

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“…To measure de novo protein biosynthesis a modified protocol described previously was used (Schopf et al, 2019;Esposito and Kinzy, 2014). Cultures of methionine prototroph R1158 (URA3::CMV-tTA MATa his3-1 leu2-0 MET15) yeast cells were grown in 5 mL CSM (-Met) over night.…”

Section: In Vivo 35 S-methionine Incorporationmentioning

confidence: 99%