Evolutionary conservation of the structure and function of meiotic Rec114-Mei4 and Mer2 complexes.
DNA double-strand break
S. cerevisiae
biomolecular condensation
meiotic recombination
multivalent protein–DNA interactions
protein structure
Journal
Genes & development
ISSN: 1549-5477
Titre abrégé: Genes Dev
Pays: United States
ID NLM: 8711660
Informations de publication
Date de publication:
01 06 2023
01 06 2023
Historique:
received:
22
01
2023
accepted:
22
06
2023
medline:
20
7
2023
pubmed:
14
7
2023
entrez:
13
7
2023
Statut:
ppublish
Résumé
Meiosis-specific Rec114-Mei4 and Mer2 complexes are thought to enable Spo11-mediated DNA double-strand break (DSB) formation through a mechanism that involves DNA-dependent condensation. However, the structure, molecular properties, and evolutionary conservation of Rec114-Mei4 and Mer2 are unclear. Here, we present AlphaFold models of Rec114-Mei4 and Mer2 complexes supported by nuclear magnetic resonance (NMR) spectroscopy, small-angle X-ray scattering (SAXS), and mutagenesis. We show that dimers composed of the Rec114 C terminus form α-helical chains that cup an N-terminal Mei4 α helix, and that Mer2 forms a parallel homotetrameric coiled coil. Both Rec114-Mei4 and Mer2 bind preferentially to branched DNA substrates, indicative of multivalent protein-DNA interactions. Indeed, the Rec114-Mei4 interaction domain contains two DNA-binding sites that point in opposite directions and drive condensation. The Mer2 coiled-coil domain bridges coaligned DNA duplexes, likely through extensive electrostatic interactions along the length of the coiled coil. Finally, we show that the structures of Rec114-Mei4 and Mer2 are conserved across eukaryotes, while DNA-binding properties vary significantly. This work provides insights into the mechanism whereby Rec114-Mei4 and Mer2 complexes promote the assembly of the meiotic DSB machinery and suggests a model in which Mer2 condensation is the essential driver of assembly, with the DNA-binding activity of Rec114-Mei4 playing a supportive role.
Identifiants
pubmed: 37442581
pii: gad.350462.123
doi: 10.1101/gad.350462.123
pmc: PMC10393190
doi:
Substances chimiques
Saccharomyces cerevisiae Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
535-553Subventions
Organisme : European Research Council
ID : 802525
Pays : International
Informations de copyright
© 2023 Daccache et al.; Published by Cold Spring Harbor Laboratory Press.
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