Dynamic Association of ORCA with Prereplicative Complex Components Regulates DNA Replication Initiation
ABSTRACT In eukaryotes, initiation of DNA replication requires the assembly of a multiprotein prereplicative complex (pre-RC) at the origins. We recently reported that a WD repeat-containing protein, origin recognition complex (ORC)-associated (ORCA/LRWD1), plays a crucial role in stabilizing ORC to...
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Online Access: | http://dx.doi.org/10.1128/mcb.00362-12 https://journals.asm.org/doi/pdf/10.1128/MCB.00362-12 |
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crasmicro:10.1128/mcb.00362-12 2023-05-15T17:52:55+02:00 Dynamic Association of ORCA with Prereplicative Complex Components Regulates DNA Replication Initiation Shen, Zhen Chakraborty, Arindam Jain, Ankur Giri, Sumanprava Ha, Taekjip Prasanth, Kannanganattu V. Prasanth, Supriya G. 2012 http://dx.doi.org/10.1128/mcb.00362-12 https://journals.asm.org/doi/pdf/10.1128/MCB.00362-12 en eng American Society for Microbiology https://journals.asm.org/non-commercial-tdm-license Molecular and Cellular Biology volume 32, issue 15, page 3107-3120 ISSN 0270-7306 1098-5549 Cell Biology Molecular Biology journal-article 2012 crasmicro https://doi.org/10.1128/mcb.00362-12 2022-10-30T09:11:21Z ABSTRACT In eukaryotes, initiation of DNA replication requires the assembly of a multiprotein prereplicative complex (pre-RC) at the origins. We recently reported that a WD repeat-containing protein, origin recognition complex (ORC)-associated (ORCA/LRWD1), plays a crucial role in stabilizing ORC to chromatin. Here, we find that ORCA is required for the G 1 -to-S-phase transition in human cells. In addition to binding to ORC, ORCA associates with Cdt1 and its inhibitor, geminin. Single-molecule pulldown experiments demonstrate that each molecule of ORCA can bind to one molecule of ORC, one molecule of Cdt1, and two molecules of geminin. Further, ORCA directly interacts with the N terminus of Orc2, and the stability of ORCA is dependent on its association with Orc2. ORCA associates with Orc2 throughout the cell cycle, with Cdt1 during mitosis and G 1 , and with geminin in post-G 1 cells. Overexpression of geminin results in the loss of interaction between ORCA and Cdt1, suggesting that increased levels of geminin in post-G 1 cells titrate Cdt1 away from ORCA. We propose that the dynamic association of ORCA with pre-RC components modulates the assembly of its interacting partners on chromatin and facilitates DNA replication initiation. Article in Journal/Newspaper Orca ASM Journals (American Society for Microbiology - via Crossref) Molecular and Cellular Biology 32 15 3107 3120 |
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ASM Journals (American Society for Microbiology - via Crossref) |
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English |
topic |
Cell Biology Molecular Biology |
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Cell Biology Molecular Biology Shen, Zhen Chakraborty, Arindam Jain, Ankur Giri, Sumanprava Ha, Taekjip Prasanth, Kannanganattu V. Prasanth, Supriya G. Dynamic Association of ORCA with Prereplicative Complex Components Regulates DNA Replication Initiation |
topic_facet |
Cell Biology Molecular Biology |
description |
ABSTRACT In eukaryotes, initiation of DNA replication requires the assembly of a multiprotein prereplicative complex (pre-RC) at the origins. We recently reported that a WD repeat-containing protein, origin recognition complex (ORC)-associated (ORCA/LRWD1), plays a crucial role in stabilizing ORC to chromatin. Here, we find that ORCA is required for the G 1 -to-S-phase transition in human cells. In addition to binding to ORC, ORCA associates with Cdt1 and its inhibitor, geminin. Single-molecule pulldown experiments demonstrate that each molecule of ORCA can bind to one molecule of ORC, one molecule of Cdt1, and two molecules of geminin. Further, ORCA directly interacts with the N terminus of Orc2, and the stability of ORCA is dependent on its association with Orc2. ORCA associates with Orc2 throughout the cell cycle, with Cdt1 during mitosis and G 1 , and with geminin in post-G 1 cells. Overexpression of geminin results in the loss of interaction between ORCA and Cdt1, suggesting that increased levels of geminin in post-G 1 cells titrate Cdt1 away from ORCA. We propose that the dynamic association of ORCA with pre-RC components modulates the assembly of its interacting partners on chromatin and facilitates DNA replication initiation. |
format |
Article in Journal/Newspaper |
author |
Shen, Zhen Chakraborty, Arindam Jain, Ankur Giri, Sumanprava Ha, Taekjip Prasanth, Kannanganattu V. Prasanth, Supriya G. |
author_facet |
Shen, Zhen Chakraborty, Arindam Jain, Ankur Giri, Sumanprava Ha, Taekjip Prasanth, Kannanganattu V. Prasanth, Supriya G. |
author_sort |
Shen, Zhen |
title |
Dynamic Association of ORCA with Prereplicative Complex Components Regulates DNA Replication Initiation |
title_short |
Dynamic Association of ORCA with Prereplicative Complex Components Regulates DNA Replication Initiation |
title_full |
Dynamic Association of ORCA with Prereplicative Complex Components Regulates DNA Replication Initiation |
title_fullStr |
Dynamic Association of ORCA with Prereplicative Complex Components Regulates DNA Replication Initiation |
title_full_unstemmed |
Dynamic Association of ORCA with Prereplicative Complex Components Regulates DNA Replication Initiation |
title_sort |
dynamic association of orca with prereplicative complex components regulates dna replication initiation |
publisher |
American Society for Microbiology |
publishDate |
2012 |
url |
http://dx.doi.org/10.1128/mcb.00362-12 https://journals.asm.org/doi/pdf/10.1128/MCB.00362-12 |
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Orca |
genre_facet |
Orca |
op_source |
Molecular and Cellular Biology volume 32, issue 15, page 3107-3120 ISSN 0270-7306 1098-5549 |
op_rights |
https://journals.asm.org/non-commercial-tdm-license |
op_doi |
https://doi.org/10.1128/mcb.00362-12 |
container_title |
Molecular and Cellular Biology |
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32 |
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15 |
container_start_page |
3107 |
op_container_end_page |
3120 |
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1766160681808166912 |