The human CTF4-orthologue AND-1 interacts with DNA polymerase α/primase via its unique C-terminal HMG box
A dynamic multi-protein assembly known as the replisome is responsible for DNA synthesis in eukaryotic cells. In yeast, the hub protein Ctf4 bridges DNA helicase and DNA polymerase and recruits factors with roles in metabolic processes coupled to DNA replication. An important question in DNA replica...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
The Royal Society
2017-01-01
|
| Series: | Open Biology |
| Subjects: | |
| Online Access: | https://royalsocietypublishing.org/doi/pdf/10.1098/rsob.170217 |
| _version_ | 1828438769957601280 |
|---|---|
| author | Mairi L. Kilkenny Aline C. Simon Jack Mainwaring David Wirthensohn Sandro Holzer Luca Pellegrini |
| author_facet | Mairi L. Kilkenny Aline C. Simon Jack Mainwaring David Wirthensohn Sandro Holzer Luca Pellegrini |
| author_sort | Mairi L. Kilkenny |
| collection | DOAJ |
| description | A dynamic multi-protein assembly known as the replisome is responsible for DNA synthesis in eukaryotic cells. In yeast, the hub protein Ctf4 bridges DNA helicase and DNA polymerase and recruits factors with roles in metabolic processes coupled to DNA replication. An important question in DNA replication is the extent to which the molecular architecture of the replisome is conserved between yeast and higher eukaryotes. Here, we describe the biochemical basis for the interaction of the human CTF4-orthologue AND-1 with DNA polymerase α (Pol α)/primase, the replicative polymerase that initiates DNA synthesis. AND-1 has maintained the trimeric structure of yeast Ctf4, driven by its conserved SepB domain. However, the primary interaction of AND-1 with Pol α/primase is mediated by its C-terminal HMG box, unique to mammalian AND-1, which binds the B subunit, at the same site targeted by the SV40 T-antigen for viral replication. In addition, we report a novel DNA-binding activity in AND-1, which might promote the correct positioning of Pol α/primase on the lagging-strand template at the replication fork. Our findings provide a biochemical basis for the specific interaction between two critical components of the human replisome, and indicate that important principles of replisome architecture have changed significantly in evolution. |
| first_indexed | 2024-12-10T20:11:45Z |
| format | Article |
| id | doaj.art-d02f8648c818478099e9295c0dede697 |
| institution | Directory Open Access Journal |
| issn | 2046-2441 |
| language | English |
| last_indexed | 2024-12-10T20:11:45Z |
| publishDate | 2017-01-01 |
| publisher | The Royal Society |
| record_format | Article |
| series | Open Biology |
| spelling | doaj.art-d02f8648c818478099e9295c0dede6972022-12-22T01:35:17ZengThe Royal SocietyOpen Biology2046-24412017-01-0171110.1098/rsob.170217170217The human CTF4-orthologue AND-1 interacts with DNA polymerase α/primase via its unique C-terminal HMG boxMairi L. KilkennyAline C. SimonJack MainwaringDavid WirthensohnSandro HolzerLuca PellegriniA dynamic multi-protein assembly known as the replisome is responsible for DNA synthesis in eukaryotic cells. In yeast, the hub protein Ctf4 bridges DNA helicase and DNA polymerase and recruits factors with roles in metabolic processes coupled to DNA replication. An important question in DNA replication is the extent to which the molecular architecture of the replisome is conserved between yeast and higher eukaryotes. Here, we describe the biochemical basis for the interaction of the human CTF4-orthologue AND-1 with DNA polymerase α (Pol α)/primase, the replicative polymerase that initiates DNA synthesis. AND-1 has maintained the trimeric structure of yeast Ctf4, driven by its conserved SepB domain. However, the primary interaction of AND-1 with Pol α/primase is mediated by its C-terminal HMG box, unique to mammalian AND-1, which binds the B subunit, at the same site targeted by the SV40 T-antigen for viral replication. In addition, we report a novel DNA-binding activity in AND-1, which might promote the correct positioning of Pol α/primase on the lagging-strand template at the replication fork. Our findings provide a biochemical basis for the specific interaction between two critical components of the human replisome, and indicate that important principles of replisome architecture have changed significantly in evolution.https://royalsocietypublishing.org/doi/pdf/10.1098/rsob.170217dna replicationreplisomeprotein–protein interactionsprotein–dna interactionsdna polymeraseprotein hub |
| spellingShingle | Mairi L. Kilkenny Aline C. Simon Jack Mainwaring David Wirthensohn Sandro Holzer Luca Pellegrini The human CTF4-orthologue AND-1 interacts with DNA polymerase α/primase via its unique C-terminal HMG box Open Biology dna replication replisome protein–protein interactions protein–dna interactions dna polymerase protein hub |
| title | The human CTF4-orthologue AND-1 interacts with DNA polymerase α/primase via its unique C-terminal HMG box |
| title_full | The human CTF4-orthologue AND-1 interacts with DNA polymerase α/primase via its unique C-terminal HMG box |
| title_fullStr | The human CTF4-orthologue AND-1 interacts with DNA polymerase α/primase via its unique C-terminal HMG box |
| title_full_unstemmed | The human CTF4-orthologue AND-1 interacts with DNA polymerase α/primase via its unique C-terminal HMG box |
| title_short | The human CTF4-orthologue AND-1 interacts with DNA polymerase α/primase via its unique C-terminal HMG box |
| title_sort | human ctf4 orthologue and 1 interacts with dna polymerase α primase via its unique c terminal hmg box |
| topic | dna replication replisome protein–protein interactions protein–dna interactions dna polymerase protein hub |
| url | https://royalsocietypublishing.org/doi/pdf/10.1098/rsob.170217 |
| work_keys_str_mv | AT mairilkilkenny thehumanctf4orthologueand1interactswithdnapolymeraseaprimaseviaitsuniquecterminalhmgbox AT alinecsimon thehumanctf4orthologueand1interactswithdnapolymeraseaprimaseviaitsuniquecterminalhmgbox AT jackmainwaring thehumanctf4orthologueand1interactswithdnapolymeraseaprimaseviaitsuniquecterminalhmgbox AT davidwirthensohn thehumanctf4orthologueand1interactswithdnapolymeraseaprimaseviaitsuniquecterminalhmgbox AT sandroholzer thehumanctf4orthologueand1interactswithdnapolymeraseaprimaseviaitsuniquecterminalhmgbox AT lucapellegrini thehumanctf4orthologueand1interactswithdnapolymeraseaprimaseviaitsuniquecterminalhmgbox AT mairilkilkenny humanctf4orthologueand1interactswithdnapolymeraseaprimaseviaitsuniquecterminalhmgbox AT alinecsimon humanctf4orthologueand1interactswithdnapolymeraseaprimaseviaitsuniquecterminalhmgbox AT jackmainwaring humanctf4orthologueand1interactswithdnapolymeraseaprimaseviaitsuniquecterminalhmgbox AT davidwirthensohn humanctf4orthologueand1interactswithdnapolymeraseaprimaseviaitsuniquecterminalhmgbox AT sandroholzer humanctf4orthologueand1interactswithdnapolymeraseaprimaseviaitsuniquecterminalhmgbox AT lucapellegrini humanctf4orthologueand1interactswithdnapolymeraseaprimaseviaitsuniquecterminalhmgbox |