Molecular choreography of primer synthesis by the eukaryotic Pol α-primase
Abstract The eukaryotic polymerase α (Pol α) synthesizes an RNA-DNA hybrid primer of 20–30 nucleotides. Pol α is composed of Pol1, Pol12, Primase 1 (Pri1), and Pri2. Pol1 and Pri1 contain the DNA polymerase and RNA primase activities, respectively. It has been unclear how Pol α hands over an RNA pri...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-06-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-39441-1 |
| _version_ | 1797795573975220224 |
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| author | Zuanning Yuan Roxana Georgescu Huilin Li Michael E. O’Donnell |
| author_facet | Zuanning Yuan Roxana Georgescu Huilin Li Michael E. O’Donnell |
| author_sort | Zuanning Yuan |
| collection | DOAJ |
| description | Abstract The eukaryotic polymerase α (Pol α) synthesizes an RNA-DNA hybrid primer of 20–30 nucleotides. Pol α is composed of Pol1, Pol12, Primase 1 (Pri1), and Pri2. Pol1 and Pri1 contain the DNA polymerase and RNA primase activities, respectively. It has been unclear how Pol α hands over an RNA primer from Pri1 to Pol1 for DNA primer extension, and how the primer length is defined. Here we report the cryo-EM analysis of yeast Pol α in the apo, primer initiation, primer elongation, RNA primer hand-off from Pri1 to Pol1, and DNA extension states, revealing a series of very large movements. We reveal a critical point at which Pol1-core moves to take over the 3’-end of the RNA from Pri1. DNA extension is limited by a spiral motion of Pol1-core. Since both Pri1 and Pol1-core are flexibly attached to a stable platform, primer growth produces stress that limits the primer length. |
| first_indexed | 2024-03-13T03:20:00Z |
| format | Article |
| id | doaj.art-4895763778d24baebc55b930840bbe07 |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-13T03:20:00Z |
| publishDate | 2023-06-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-4895763778d24baebc55b930840bbe072023-06-25T11:22:12ZengNature PortfolioNature Communications2041-17232023-06-0114111410.1038/s41467-023-39441-1Molecular choreography of primer synthesis by the eukaryotic Pol α-primaseZuanning Yuan0Roxana Georgescu1Huilin Li2Michael E. O’Donnell3Department of Structural Biology, Van Andel InstituteDNA Replication Laboratory and Howard Hughes Medical Institute, Rockefeller UniversityDepartment of Structural Biology, Van Andel InstituteDNA Replication Laboratory and Howard Hughes Medical Institute, Rockefeller UniversityAbstract The eukaryotic polymerase α (Pol α) synthesizes an RNA-DNA hybrid primer of 20–30 nucleotides. Pol α is composed of Pol1, Pol12, Primase 1 (Pri1), and Pri2. Pol1 and Pri1 contain the DNA polymerase and RNA primase activities, respectively. It has been unclear how Pol α hands over an RNA primer from Pri1 to Pol1 for DNA primer extension, and how the primer length is defined. Here we report the cryo-EM analysis of yeast Pol α in the apo, primer initiation, primer elongation, RNA primer hand-off from Pri1 to Pol1, and DNA extension states, revealing a series of very large movements. We reveal a critical point at which Pol1-core moves to take over the 3’-end of the RNA from Pri1. DNA extension is limited by a spiral motion of Pol1-core. Since both Pri1 and Pol1-core are flexibly attached to a stable platform, primer growth produces stress that limits the primer length.https://doi.org/10.1038/s41467-023-39441-1 |
| spellingShingle | Zuanning Yuan Roxana Georgescu Huilin Li Michael E. O’Donnell Molecular choreography of primer synthesis by the eukaryotic Pol α-primase Nature Communications |
| title | Molecular choreography of primer synthesis by the eukaryotic Pol α-primase |
| title_full | Molecular choreography of primer synthesis by the eukaryotic Pol α-primase |
| title_fullStr | Molecular choreography of primer synthesis by the eukaryotic Pol α-primase |
| title_full_unstemmed | Molecular choreography of primer synthesis by the eukaryotic Pol α-primase |
| title_short | Molecular choreography of primer synthesis by the eukaryotic Pol α-primase |
| title_sort | molecular choreography of primer synthesis by the eukaryotic pol α primase |
| url | https://doi.org/10.1038/s41467-023-39441-1 |
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