Reconstitution of a eukaryotic replisome reveals suppression mechanisms that define leading/lagging strand operation
We have reconstituted a eukaryotic leading/lagging strand replisome comprising 31 distinct polypeptides. This study identifies a process unprecedented in bacterial replisomes. While bacteria and phage simply recruit polymerases to the fork, we find that suppression mechanisms are used to position th...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2015-04-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/04988 |
| _version_ | 1811253315948773376 |
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| author | Roxana E Georgescu Grant D Schauer Nina Y Yao Lance D Langston Olga Yurieva Dan Zhang Jeff Finkelstein Mike E O'Donnell |
| author_facet | Roxana E Georgescu Grant D Schauer Nina Y Yao Lance D Langston Olga Yurieva Dan Zhang Jeff Finkelstein Mike E O'Donnell |
| author_sort | Roxana E Georgescu |
| collection | DOAJ |
| description | We have reconstituted a eukaryotic leading/lagging strand replisome comprising 31 distinct polypeptides. This study identifies a process unprecedented in bacterial replisomes. While bacteria and phage simply recruit polymerases to the fork, we find that suppression mechanisms are used to position the distinct eukaryotic polymerases on their respective strands. Hence, Pol ε is active with CMG on the leading strand, but it is unable to function on the lagging strand, even when Pol δ is not present. Conversely, Pol δ-PCNA is the only enzyme capable of extending Okazaki fragments in the presence of Pols ε and α. We have shown earlier that Pol δ-PCNA is suppressed on the leading strand with CMG (Georgescu et al., 2014). We propose that CMG, the 11-subunit helicase, is responsible for one or both of these suppression mechanisms that spatially control polymerase occupancy at the fork. |
| first_indexed | 2024-04-12T16:48:59Z |
| format | Article |
| id | doaj.art-82749394aac14281ad3f71f0c1367891 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:48:59Z |
| publishDate | 2015-04-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-82749394aac14281ad3f71f0c13678912022-12-22T03:24:28ZengeLife Sciences Publications LtdeLife2050-084X2015-04-01410.7554/eLife.04988Reconstitution of a eukaryotic replisome reveals suppression mechanisms that define leading/lagging strand operationRoxana E Georgescu0https://orcid.org/0000-0002-1882-2358Grant D Schauer1Nina Y Yao2Lance D Langston3https://orcid.org/0000-0002-2736-9284Olga Yurieva4Dan Zhang5Jeff Finkelstein6Mike E O'Donnell7DNA Replication Laboratory, Howard Hughes Medical Institute, Rockefeller University, New York, United StatesDNA Replication Laboratory, Howard Hughes Medical Institute, Rockefeller University, New York, United StatesDNA Replication Laboratory, Howard Hughes Medical Institute, Rockefeller University, New York, United StatesDNA Replication Laboratory, Howard Hughes Medical Institute, Rockefeller University, New York, United StatesDNA Replication Laboratory, Howard Hughes Medical Institute, Rockefeller University, New York, United StatesDNA Replication Laboratory, Howard Hughes Medical Institute, Rockefeller University, New York, United StatesDNA Replication Laboratory, Howard Hughes Medical Institute, Rockefeller University, New York, United StatesDNA Replication Laboratory, Howard Hughes Medical Institute, Rockefeller University, New York, United StatesWe have reconstituted a eukaryotic leading/lagging strand replisome comprising 31 distinct polypeptides. This study identifies a process unprecedented in bacterial replisomes. While bacteria and phage simply recruit polymerases to the fork, we find that suppression mechanisms are used to position the distinct eukaryotic polymerases on their respective strands. Hence, Pol ε is active with CMG on the leading strand, but it is unable to function on the lagging strand, even when Pol δ is not present. Conversely, Pol δ-PCNA is the only enzyme capable of extending Okazaki fragments in the presence of Pols ε and α. We have shown earlier that Pol δ-PCNA is suppressed on the leading strand with CMG (Georgescu et al., 2014). We propose that CMG, the 11-subunit helicase, is responsible for one or both of these suppression mechanisms that spatially control polymerase occupancy at the fork.https://elifesciences.org/articles/04988DNA replicationreplication forkCMGPol deltaPol epsilon |
| spellingShingle | Roxana E Georgescu Grant D Schauer Nina Y Yao Lance D Langston Olga Yurieva Dan Zhang Jeff Finkelstein Mike E O'Donnell Reconstitution of a eukaryotic replisome reveals suppression mechanisms that define leading/lagging strand operation eLife DNA replication replication fork CMG Pol delta Pol epsilon |
| title | Reconstitution of a eukaryotic replisome reveals suppression mechanisms that define leading/lagging strand operation |
| title_full | Reconstitution of a eukaryotic replisome reveals suppression mechanisms that define leading/lagging strand operation |
| title_fullStr | Reconstitution of a eukaryotic replisome reveals suppression mechanisms that define leading/lagging strand operation |
| title_full_unstemmed | Reconstitution of a eukaryotic replisome reveals suppression mechanisms that define leading/lagging strand operation |
| title_short | Reconstitution of a eukaryotic replisome reveals suppression mechanisms that define leading/lagging strand operation |
| title_sort | reconstitution of a eukaryotic replisome reveals suppression mechanisms that define leading lagging strand operation |
| topic | DNA replication replication fork CMG Pol delta Pol epsilon |
| url | https://elifesciences.org/articles/04988 |
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