Structural and mutational analysis of the ribosome-arresting human XBP1u
XBP1u, a central component of the unfolded protein response (UPR), is a mammalian protein containing a functionally critical translational arrest peptide (AP). Here, we present a 3 Å cryo-EM structure of the stalled human XBP1u AP. It forms a unique turn in the ribosomal exit tunnel proximal to the...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2019-06-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/46267 |
| _version_ | 1818026962537414656 |
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| author | Vivekanandan Shanmuganathan Nina Schiller Anastasia Magoulopoulou Jingdong Cheng Katharina Braunger Florian Cymer Otto Berninghausen Birgitta Beatrix Kenji Kohno Gunnar von Heijne Roland Beckmann |
| author_facet | Vivekanandan Shanmuganathan Nina Schiller Anastasia Magoulopoulou Jingdong Cheng Katharina Braunger Florian Cymer Otto Berninghausen Birgitta Beatrix Kenji Kohno Gunnar von Heijne Roland Beckmann |
| author_sort | Vivekanandan Shanmuganathan |
| collection | DOAJ |
| description | XBP1u, a central component of the unfolded protein response (UPR), is a mammalian protein containing a functionally critical translational arrest peptide (AP). Here, we present a 3 Å cryo-EM structure of the stalled human XBP1u AP. It forms a unique turn in the ribosomal exit tunnel proximal to the peptidyl transferase center where it causes a subtle distortion, thereby explaining the temporary translational arrest induced by XBP1u. During ribosomal pausing the hydrophobic region 2 (HR2) of XBP1u is recognized by SRP, but fails to efficiently gate the Sec61 translocon. An exhaustive mutagenesis scan of the XBP1u AP revealed that only 8 out of 20 mutagenized positions are optimal; in the remaining 12 positions, we identify 55 different mutations increase the level of translational arrest. Thus, the wildtype XBP1u AP induces only an intermediate level of translational arrest, allowing efficient targeting by SRP without activating the Sec61 channel. |
| first_indexed | 2024-12-10T04:40:20Z |
| format | Article |
| id | doaj.art-f441759e29854117a41889883f5b50a1 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-12-10T04:40:20Z |
| publishDate | 2019-06-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-f441759e29854117a41889883f5b50a12022-12-22T02:01:55ZengeLife Sciences Publications LtdeLife2050-084X2019-06-01810.7554/eLife.46267Structural and mutational analysis of the ribosome-arresting human XBP1uVivekanandan Shanmuganathan0Nina Schiller1Anastasia Magoulopoulou2Jingdong Cheng3Katharina Braunger4https://orcid.org/0000-0002-9067-2155Florian Cymer5Otto Berninghausen6Birgitta Beatrix7Kenji Kohno8https://orcid.org/0000-0002-3503-6551Gunnar von Heijne9https://orcid.org/0000-0002-4490-8569Roland Beckmann10https://orcid.org/0000-0003-4291-3898Gene Center, Department of Biochemistry, Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Munich, GermanyDepartment of Biochemistry and Biophysics, Stockholm University, Stockholm, SwedenDepartment of Biochemistry and Biophysics, Stockholm University, Stockholm, SwedenGene Center, Department of Biochemistry, Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Munich, GermanyGene Center, Department of Biochemistry, Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Munich, GermanyDepartment of Biochemistry and Biophysics, Stockholm University, Stockholm, SwedenGene Center, Department of Biochemistry, Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Munich, GermanyGene Center, Department of Biochemistry, Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Munich, GermanyInstitute for Research Initiatives, Nara Institute of Science and Technology, Takayama, JapanDepartment of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden; Science for Life Laboratory, Stockholm University, Solna, SwedenGene Center, Department of Biochemistry, Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Munich, GermanyXBP1u, a central component of the unfolded protein response (UPR), is a mammalian protein containing a functionally critical translational arrest peptide (AP). Here, we present a 3 Å cryo-EM structure of the stalled human XBP1u AP. It forms a unique turn in the ribosomal exit tunnel proximal to the peptidyl transferase center where it causes a subtle distortion, thereby explaining the temporary translational arrest induced by XBP1u. During ribosomal pausing the hydrophobic region 2 (HR2) of XBP1u is recognized by SRP, but fails to efficiently gate the Sec61 translocon. An exhaustive mutagenesis scan of the XBP1u AP revealed that only 8 out of 20 mutagenized positions are optimal; in the remaining 12 positions, we identify 55 different mutations increase the level of translational arrest. Thus, the wildtype XBP1u AP induces only an intermediate level of translational arrest, allowing efficient targeting by SRP without activating the Sec61 channel.https://elifesciences.org/articles/46267translational pausingribosomeXBP1 |
| spellingShingle | Vivekanandan Shanmuganathan Nina Schiller Anastasia Magoulopoulou Jingdong Cheng Katharina Braunger Florian Cymer Otto Berninghausen Birgitta Beatrix Kenji Kohno Gunnar von Heijne Roland Beckmann Structural and mutational analysis of the ribosome-arresting human XBP1u eLife translational pausing ribosome XBP1 |
| title | Structural and mutational analysis of the ribosome-arresting human XBP1u |
| title_full | Structural and mutational analysis of the ribosome-arresting human XBP1u |
| title_fullStr | Structural and mutational analysis of the ribosome-arresting human XBP1u |
| title_full_unstemmed | Structural and mutational analysis of the ribosome-arresting human XBP1u |
| title_short | Structural and mutational analysis of the ribosome-arresting human XBP1u |
| title_sort | structural and mutational analysis of the ribosome arresting human xbp1u |
| topic | translational pausing ribosome XBP1 |
| url | https://elifesciences.org/articles/46267 |
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