Kadlec J.,European Molecular Biology Laboratory |
Kadlec J.,Joseph Fourier University |
Kadlec J.,Diamond Light Source |
Hallacli E.,European Molecular Biology Laboratory |
And 12 more authors.
Nature Structural and Molecular Biology | Year: 2011
The male-specific lethal (MSL) complex is required for dosage compensation in Drosophila melanogaster, and analogous complexes exist in mammals. We report structures of binary complexes of mammalian MSL3 and the histone acetyltransferase (HAT) MOF with consecutive segments of MSL1. MSL1 interacts with MSL3 as an extended chain forming an extensive hydrophobic interface, whereas the MSL1-MOF interface involves electrostatic interactions between the HAT domain and a long helix of MSL1. This structure provides insights into the catalytic mechanism of MOF and enables us to show analogous interactions of MOF with NSL1. In Drosophila, selective disruption of Msl1 interactions with Msl3 or Mof severely affects Msl1 targeting to the body of dosage-compensated genes and several high-affinity sites, without affecting promoter binding. We propose that Msl1 acts as a scaffold for MSL complex assembly to achieve specific targeting to the X chromosome. © 2011 Nature America, Inc. All rights reserved.
Hallacli E.,Max Planck Institute For Immunbiologie Und Epigenetik |
Lipp M.,European Molecular Biology Laboratory |
Lipp M.,French National Center for Scientific Research |
Georgiev P.,Max Planck Institute For Immunbiologie Und Epigenetik |
And 6 more authors.
Molecular Cell | Year: 2012
The Male-Specific Lethal (MSL) complex regulates dosage compensation of the male X chromosome in Drosophila. Here, we report the crystal structure of its MSL1/MSL2 core, where two MSL2 subunits bind to a dimer formed by two molecules of MSL1. Analysis of structure-based mutants revealed that MSL2 can only interact with the MSL1 dimer, but MSL1 dimerization is MSL2 independent. We show that Msl1 is a substrate for Msl2 E3 ubiquitin ligase activity. ChIP experiments revealed that Msl1 dimerization is essential for targeting and spreading of the MSL complex on X-linked genes; however, Msl1 binding to promoters of male and female cells is independent of the dimer status and other MSL proteins. Finally, we show that loss of Msl1 dimerization leads to male-specific lethality. We propose that Msl1-mediated dimerization of the entire MSL complex is required for Msl2 binding, X chromosome recognition, and spreading along the X chromosome. © 2012 Elsevier Inc.
Bonisch U.,Max Planck Institute For Immunbiologie Und Epigenetik |
Arrigoni L.,Max Planck Institute For Immunbiologie Und Epigenetik |
Betancourt E.,Max Planck Institute For Immunbiologie Und Epigenetik |
Bruder K.,Max Planck Institute For Immunbiologie Und Epigenetik
BioSpektrum | Year: 2015
Manual library preparation for ChIP-Seq applications is very tedious to conduct and lacks process standardization. Here we translate a commercially available ChIP-Seq library preparation protocol into commands for the Eppendorf epMotion automated liquid handling system and show that libraries with comparable quantity and quality can be easily generated, liberating lab personnel from repetitive processing steps while maximizing reproducibility and data comparability. © 2015, Springer-Verlag Berlin Heidelberg.