TY - JOUR
T1 - Novel mechanistic insights into the role of Mer2 as the keystone of meiotic DNA break formation
AU - Rousova, Dorota
AU - Nivsarkar, Vaishnavi
AU - Altmannova, Veronika
AU - Raina, Vivek B.
AU - Funk, Saskia K.
AU - Liedtke, David
AU - Janning, Petra
AU - Müller, Franziska
AU - Reichle, Heidi
AU - Vader, Gerben
AU - Weir, John R.
N1 - Funding Information:
We are extremely grateful to Francesca Mattiroli (Hubrecht Institute, Utrecht), for assistance with producing mononucleosomes. The histone H3 expression plasmid pET3 and the plasmid pUC19_601-147 DNA were a kind gift from Francesca Mattiroli. We are also grateful to Luke Berchowitz (Columbia University, New York), Andreas Hochwagen (NYU, New York) and Eric Greene (Columbia University, New York) for support, reagents and expertise. The plasmid pUC18_601-167 DNA was a kind gift from Andrea Musacchio (MPI of Molecular Physiology, Dortmund). We thank Christopher Heim (MPI Developmental Biology) for technical help with SEC-MALS and MST measurements. We thank Andreas Blaha and Constanze Gremmelmaier for their technical support. Quantitative Mass-Spec and analysis was made possible by the EMBL Proteomics Core Facility (EMBL, Heidelberg). Work in the Weir lab is funded by the Max Planck Society and DFG grant WE 6513/2-1. Work in the Vader lab was funded by Max Planck Society and the European Research Council (ERC StG URDNA; agreement no. 638197), and an Amsterdam UMC Research Fellowship. SKF is funded by a studentship from the International Max Planck Research School (IMPRS) ?From Molecules to Organisms?.
Funding Information:
and Eric Greene (Columbia University, New York) for support, reagents and expertise. The plasmid pUC18_601-167 DNA was a kind gift from Andrea Musacchio (MPI of Molecular Physiology, Dortmund). We thank Christopher Heim (MPI Developmental Biology) for technical help with SEC-MALS and MST measurements. We thank Andreas Blaha and Constanze Gremmelmaier for their technical support. Quantitative Mass-Spec and analysis was made possible by the EMBL Proteomics Core Facility (EMBL, Heidelberg). Work in the Weir lab is funded by the Max Planck Society and DFG grant WE 6513/2-1. Work in the Vader lab was funded by Max Planck Society and the European Research Council (ERC StG URDNA; agreement no. 638197), and an Amsterdam UMC Research Fellowship. SKF is funded by a studentship from the International Max Planck Research School (IMPRS) “From Molecules to Organisms”.
Publisher Copyright:
© 2021, eLife Sciences Publications Ltd. All rights reserved.
PY - 2021/12/1
Y1 - 2021/12/1
N2 - In meiosis, DNA double strand break (DSB) formation by Spo11 initiates recombination and enables chromosome segregation. Numerous factors are required for Spo11 activity, and couple the DSB machinery to the development of a meiosis-specific “axis-tethered loop” chromosome organization. Through in vitro reconstitution and budding yeast genetics we here provide architectural insight into the DSB machinery by focussing on a foundational DSB factor, Mer2. We characterise the interaction of Mer2 with the histone reader Spp1, and show that Mer2 directly associates to nucleosomes, likely highlighting a contribution of Mer2 to tethering DSB factors to chromatin. We reveal the biochemical basis of Mer2 association with Hop1, a HORMA domain-containing chromosomal axis factor. Finally, we identify a conserved region within Mer2 crucial for DSB activity, and show that this region of Mer2 interacts with the DSB factor Mre11. In combination with previous work, we establish Mer2 as a keystone of the DSB machinery by bridging key protein complexes involved in the initiation of meiotic recombination.
AB - In meiosis, DNA double strand break (DSB) formation by Spo11 initiates recombination and enables chromosome segregation. Numerous factors are required for Spo11 activity, and couple the DSB machinery to the development of a meiosis-specific “axis-tethered loop” chromosome organization. Through in vitro reconstitution and budding yeast genetics we here provide architectural insight into the DSB machinery by focussing on a foundational DSB factor, Mer2. We characterise the interaction of Mer2 with the histone reader Spp1, and show that Mer2 directly associates to nucleosomes, likely highlighting a contribution of Mer2 to tethering DSB factors to chromatin. We reveal the biochemical basis of Mer2 association with Hop1, a HORMA domain-containing chromosomal axis factor. Finally, we identify a conserved region within Mer2 crucial for DSB activity, and show that this region of Mer2 interacts with the DSB factor Mre11. In combination with previous work, we establish Mer2 as a keystone of the DSB machinery by bridging key protein complexes involved in the initiation of meiotic recombination.
UR - https://www.scopus.com/pages/publications/85122008507
U2 - 10.7554/eLife.72330
DO - 10.7554/eLife.72330
M3 - Article
C2 - 34951404
SN - 2050-084X
VL - 10
JO - eLife
JF - eLife
M1 - e72330
ER -