A new class of disordered elements controls DNA replication through initiator self-assembly
The initiation of DNA replication in metazoans occurs at thousands of chromosomal sites known as origins. At each origin, the Origin Recognition Complex (ORC), Cdc6, and Cdt1 co-assemble to load the Mcm2-7 replicative helicase onto chromatin. Current replication models envisage a linear arrangement...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2019-09-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/48562 |
| _version_ | 1818023870979899392 |
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| author | Matthew W Parker Maren Bell Mustafa Mir Jonchee A Kao Xavier Darzacq Michael R Botchan James M Berger |
| author_facet | Matthew W Parker Maren Bell Mustafa Mir Jonchee A Kao Xavier Darzacq Michael R Botchan James M Berger |
| author_sort | Matthew W Parker |
| collection | DOAJ |
| description | The initiation of DNA replication in metazoans occurs at thousands of chromosomal sites known as origins. At each origin, the Origin Recognition Complex (ORC), Cdc6, and Cdt1 co-assemble to load the Mcm2-7 replicative helicase onto chromatin. Current replication models envisage a linear arrangement of isolated origins functioning autonomously; the extent of inter-origin organization and communication is unknown. Here, we report that the replication initiation machinery of D. melanogaster unexpectedly undergoes liquid-liquid phase separation (LLPS) upon binding DNA in vitro. We find that ORC, Cdc6, and Cdt1 contain intrinsically disordered regions (IDRs) that drive LLPS and constitute a new class of phase separating elements. Initiator IDRs are shown to regulate multiple functions, including chromosome recruitment, initiator-specific co-assembly, and Mcm2-7 loading. These data help explain how CDK activity controls replication initiation and suggest that replication programs are subject to higher-order levels of inter-origin organization. |
| first_indexed | 2024-12-10T03:51:12Z |
| format | Article |
| id | doaj.art-3b730dc3b725431c82d085509fb2026f |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-12-10T03:51:12Z |
| publishDate | 2019-09-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-3b730dc3b725431c82d085509fb2026f2022-12-22T02:03:16ZengeLife Sciences Publications LtdeLife2050-084X2019-09-01810.7554/eLife.48562A new class of disordered elements controls DNA replication through initiator self-assemblyMatthew W Parker0https://orcid.org/0000-0002-7571-0010Maren Bell1Mustafa Mir2https://orcid.org/0000-0001-8280-2821Jonchee A Kao3Xavier Darzacq4https://orcid.org/0000-0003-2537-8395Michael R Botchan5https://orcid.org/0000-0003-0459-5518James M Berger6https://orcid.org/0000-0003-0666-1240Department of Biophysics and Biophysical Chemistry, Johns Hopkins School of Medicine, Baltimore, United States; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United StatesDepartment of Biophysics and Biophysical Chemistry, Johns Hopkins School of Medicine, Baltimore, United StatesThe initiation of DNA replication in metazoans occurs at thousands of chromosomal sites known as origins. At each origin, the Origin Recognition Complex (ORC), Cdc6, and Cdt1 co-assemble to load the Mcm2-7 replicative helicase onto chromatin. Current replication models envisage a linear arrangement of isolated origins functioning autonomously; the extent of inter-origin organization and communication is unknown. Here, we report that the replication initiation machinery of D. melanogaster unexpectedly undergoes liquid-liquid phase separation (LLPS) upon binding DNA in vitro. We find that ORC, Cdc6, and Cdt1 contain intrinsically disordered regions (IDRs) that drive LLPS and constitute a new class of phase separating elements. Initiator IDRs are shown to regulate multiple functions, including chromosome recruitment, initiator-specific co-assembly, and Mcm2-7 loading. These data help explain how CDK activity controls replication initiation and suggest that replication programs are subject to higher-order levels of inter-origin organization.https://elifesciences.org/articles/48562DNA replicationphase separationintrinsically disorderedORCCdc6Cdt1 |
| spellingShingle | Matthew W Parker Maren Bell Mustafa Mir Jonchee A Kao Xavier Darzacq Michael R Botchan James M Berger A new class of disordered elements controls DNA replication through initiator self-assembly eLife DNA replication phase separation intrinsically disordered ORC Cdc6 Cdt1 |
| title | A new class of disordered elements controls DNA replication through initiator self-assembly |
| title_full | A new class of disordered elements controls DNA replication through initiator self-assembly |
| title_fullStr | A new class of disordered elements controls DNA replication through initiator self-assembly |
| title_full_unstemmed | A new class of disordered elements controls DNA replication through initiator self-assembly |
| title_short | A new class of disordered elements controls DNA replication through initiator self-assembly |
| title_sort | new class of disordered elements controls dna replication through initiator self assembly |
| topic | DNA replication phase separation intrinsically disordered ORC Cdc6 Cdt1 |
| url | https://elifesciences.org/articles/48562 |
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