A ribosome-associating chaperone mediates GTP-driven vectorial folding of nascent eEF1A
Abstract Eukaryotic translation elongation factor 1A (eEF1A) is a highly abundant, multi-domain GTPase. Post-translational steps essential for eEF1A biogenesis are carried out by bespoke chaperones but co-translational mechanisms tailored to eEF1A folding remain unexplored. Here, we use AlphaPulldow...
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Nature Portfolio
2025-02-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-56489-3 |
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| author | Ibrahim M. Sabbarini Dvir Reif Kibum Park Alexander J. McQuown Anjali R. Nelliat Charlotte Trejtnar Volker Dötsch Eugene I. Shakhnovich Andrew W. Murray Vladimir Denic |
| author_facet | Ibrahim M. Sabbarini Dvir Reif Kibum Park Alexander J. McQuown Anjali R. Nelliat Charlotte Trejtnar Volker Dötsch Eugene I. Shakhnovich Andrew W. Murray Vladimir Denic |
| author_sort | Ibrahim M. Sabbarini |
| collection | DOAJ |
| description | Abstract Eukaryotic translation elongation factor 1A (eEF1A) is a highly abundant, multi-domain GTPase. Post-translational steps essential for eEF1A biogenesis are carried out by bespoke chaperones but co-translational mechanisms tailored to eEF1A folding remain unexplored. Here, we use AlphaPulldown to identify Ypl225w (also known as Chp1, Chaperone 1 for eEF1A) as a conserved yeast protein predicted to stabilize the N-terminal, GTP-binding (G) domain of eEF1A against its misfolding propensity, as predicted by computational simulations and validated by microscopy analysis of ypl225wΔ cells. Proteomics and biochemical reconstitution reveal that Ypl225w functions as a co-translational chaperone by forming dual interactions with the eEF1A G domain nascent chain and the UBA domain of ribosome-bound nascent polypeptide-associated complex (NAC). Lastly, we show that Ypl225w primes eEF1A nascent chains for binding to GTP as part of a folding mechanism tightly coupled to chaperone recycling. Our work shows that an ATP-independent chaperone can drive vectorial folding of nascent chains by co-opting G protein nucleotide binding. |
| format | Article |
| id | doaj-art-e671cb47210f411e88fe724a443368cf |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-e671cb47210f411e88fe724a443368cf2025-02-09T12:44:35ZengNature PortfolioNature Communications2041-17232025-02-0116111810.1038/s41467-025-56489-3A ribosome-associating chaperone mediates GTP-driven vectorial folding of nascent eEF1AIbrahim M. Sabbarini0Dvir Reif1Kibum Park2Alexander J. McQuown3Anjali R. Nelliat4Charlotte Trejtnar5Volker Dötsch6Eugene I. Shakhnovich7Andrew W. Murray8Vladimir Denic9Department of Molecular and Cellular Biology, Harvard UniversityDepartment of Molecular and Cellular Biology, Harvard UniversityDepartment of Chemistry and Chemical Biology, Harvard UniversityDepartment of Molecular and Cellular Biology, Harvard UniversityGraduate Program in Systems Biology, Harvard Medical SchoolInstitute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance, Goethe UniversityInstitute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance, Goethe UniversityDepartment of Chemistry and Chemical Biology, Harvard UniversityDepartment of Molecular and Cellular Biology, Harvard UniversityDepartment of Molecular and Cellular Biology, Harvard UniversityAbstract Eukaryotic translation elongation factor 1A (eEF1A) is a highly abundant, multi-domain GTPase. Post-translational steps essential for eEF1A biogenesis are carried out by bespoke chaperones but co-translational mechanisms tailored to eEF1A folding remain unexplored. Here, we use AlphaPulldown to identify Ypl225w (also known as Chp1, Chaperone 1 for eEF1A) as a conserved yeast protein predicted to stabilize the N-terminal, GTP-binding (G) domain of eEF1A against its misfolding propensity, as predicted by computational simulations and validated by microscopy analysis of ypl225wΔ cells. Proteomics and biochemical reconstitution reveal that Ypl225w functions as a co-translational chaperone by forming dual interactions with the eEF1A G domain nascent chain and the UBA domain of ribosome-bound nascent polypeptide-associated complex (NAC). Lastly, we show that Ypl225w primes eEF1A nascent chains for binding to GTP as part of a folding mechanism tightly coupled to chaperone recycling. Our work shows that an ATP-independent chaperone can drive vectorial folding of nascent chains by co-opting G protein nucleotide binding.https://doi.org/10.1038/s41467-025-56489-3 |
| spellingShingle | Ibrahim M. Sabbarini Dvir Reif Kibum Park Alexander J. McQuown Anjali R. Nelliat Charlotte Trejtnar Volker Dötsch Eugene I. Shakhnovich Andrew W. Murray Vladimir Denic A ribosome-associating chaperone mediates GTP-driven vectorial folding of nascent eEF1A Nature Communications |
| title | A ribosome-associating chaperone mediates GTP-driven vectorial folding of nascent eEF1A |
| title_full | A ribosome-associating chaperone mediates GTP-driven vectorial folding of nascent eEF1A |
| title_fullStr | A ribosome-associating chaperone mediates GTP-driven vectorial folding of nascent eEF1A |
| title_full_unstemmed | A ribosome-associating chaperone mediates GTP-driven vectorial folding of nascent eEF1A |
| title_short | A ribosome-associating chaperone mediates GTP-driven vectorial folding of nascent eEF1A |
| title_sort | ribosome associating chaperone mediates gtp driven vectorial folding of nascent eef1a |
| url | https://doi.org/10.1038/s41467-025-56489-3 |
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