The EMBO journal, 2021-12, Vol.40 (23), p.e108819-n/a
2021
Details
- Autor(en) / Beteiligte
- Titel
- Structure of a human replisome shows the organisation and interactions of a DNA replication machine
- Ist Teil von
- The EMBO journal, 2021-12, Vol.40 (23), p.e108819-n/a
- Ort / Verlag
- England: Blackwell Publishing Ltd
- Erscheinungsjahr
- 2021
- Link zum Volltext
- Quelle
- Wiley-Blackwell Journals
- Beschreibungen/Notizen
- The human replisome is an elaborate arrangement of molecular machines responsible for accurate chromosome replication. At its heart is the CDC45‐MCM‐GINS (CMG) helicase, which, in addition to unwinding the parental DNA duplex, arranges many proteins including the leading‐strand polymerase Pol ε, together with TIMELESS‐TIPIN, CLASPIN and AND‐1 that have key and varied roles in maintaining smooth replisome progression. How these proteins are coordinated in the human replisome is poorly understood. We have determined a 3.2 Å cryo‐EM structure of a human replisome comprising CMG, Pol ε, TIMELESS‐TIPIN, CLASPIN and AND‐1 bound to replication fork DNA. The structure permits a detailed understanding of how AND‐1, TIMELESS‐TIPIN and Pol ε engage CMG, reveals how CLASPIN binds to multiple replisome components and identifies the position of the Pol ε catalytic domain. Furthermore, the intricate network of contacts contributed by MCM subunits and TIMELESS‐TIPIN with replication fork DNA suggests a mechanism for strand separation. SYNOPSIS Cryo‐EM structures of the core human replisome reveal its complex architecture and show how TIMELESS‐TIPIN, AND‐1, Pol ε and CLASPIN engage the CMG helicase. Contacts between MCM subunits and DNA suggest a mechanism for strand separation. High‐resolution structure of a reconstituted human replisome comprising the CMG helicase, AND‐1, TIMELESS‐TIPIN, Pol ε, CLASPIN and fork DNA. Detailed description of the protein‐protein and protein‐DNA contacts that underpin the organisation of the human replisome. Atomic model for three regions of CLASPIN showing how the protein extends across one side of the replisome contacting TIMELESS, MCM2 and MCM6. Identification of a specific conformation for the catalytic domain of Pol ε, demonstrating that Pol ε can adopt a “linear” configuration in the human replisome. Cryo‐EM structures of reconstituted human core replisomes show how TIMELESS‐TIPIN, AND‐1, Pol ε and CLASPIN engage the CMG helicase, and suggest a mechanism for DNA strand separation.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0261-4189eISSN: 1460-2075DOI: 10.15252/embj.2021108819Titel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_8634136
- Format
- –
- Schlagworte
- Adaptor Proteins, Signal Transducing - chemistry, Adaptor Proteins, Signal Transducing - genetics, Adaptor Proteins, Signal Transducing - metabolism, Cdc45 protein, Cell Cycle Proteins - chemistry, Cell Cycle Proteins - genetics, Cell Cycle Proteins - metabolism, Chromosomes, CMG helicase, cryo‐EM, Cytoskeletal Proteins - chemistry, Cytoskeletal Proteins - genetics, Cytoskeletal Proteins - metabolism, Deoxyribonucleic acid, DNA, DNA biosynthesis, DNA helicase, DNA Polymerase II - chemistry, DNA Polymerase II - genetics, DNA Polymerase II - metabolism, DNA Replication, DNA structure, DNA-Binding Proteins - chemistry, DNA-Binding Proteins - genetics, DNA-Binding Proteins - metabolism, Domains, fork protection complex, Humans, Intracellular Signaling Peptides and Proteins - chemistry, Intracellular Signaling Peptides and Proteins - genetics, Intracellular Signaling Peptides and Proteins - metabolism, Models, Molecular, Molecular machines, Poly-ADP-Ribose Binding Proteins - chemistry, Poly-ADP-Ribose Binding Proteins - genetics, Poly-ADP-Ribose Binding Proteins - metabolism, Protein Conformation, Protein structure, Proteins, Replication, replisome, Separation, Unwinding