Regulated Eukaryotic DNA Replication Origin Firing with Purified Proteins
| Autor: | Tom D Deegan, John F.X. Diffley, Anne Early, Joseph T.P. Yeeles, Agnieszka Janska |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
DNA Replication
Saccharomyces cerevisiae Proteins DNA replication initiation Eukaryotic DNA replication Cell Cycle Proteins Replication Origin DNA-Directed DNA Polymerase Saccharomyces cerevisiae Biology Protein Serine-Threonine Kinases Pre-replication complex Article 03 medical and health sciences 0302 clinical medicine Replication factor C Control of chromosome duplication Minichromosome maintenance Multienzyme Complexes Replication Protein A Phosphorylation 030304 developmental biology 0303 health sciences Multidisciplinary Minichromosome Maintenance Proteins Nuclear Proteins Cyclin-Dependent Kinases DNA-Binding Proteins Licensing factor Biochemistry Multiprotein Complexes Origin recognition complex 030217 neurology & neurosurgery |
| Zdroj: | Nature |
| ISSN: | 1476-4687 0028-0836 |
| Popis: | Eukaryotic cells initiate DNA replication from multiple origins, which must be tightly regulated to promote precise genome duplication in every cell cycle. To accomplish this, initiation is partitioned into two temporally discrete steps: a double hexameric minichromosome maintenance (MCM) complex is first loaded at replication origins during G1 phase, and then converted to the active CMG (Cdc45-MCM-GINS) helicase during S phase. Here we describe the reconstitution of budding yeast DNA replication initiation with 16 purified replication factors, made from 42 polypeptides. Origin-dependent initiation recapitulates regulation seen in vivo. Cyclin-dependent kinase (CDK) inhibits MCM loading by phosphorylating the origin recognition complex (ORC) and promotes CMG formation by phosphorylating Sld2 and Sld3. Dbf4-dependent kinase (DDK) promotes replication by phosphorylating MCM, and can act either before or after CDK. These experiments define the minimum complement of proteins, protein kinase substrates and co-factors required for regulated eukaryotic DNA replication. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|