(Gene Center and Department of Biochemistry. LMU Munich)
(Institut Germans Trias i Pujol. Institut de Recerca contra la Leucèmia Josep Carreras)
(Biomedical Center (BMC). Department of Physiological Chemistry. Faculty of Medicine. LMU Munich)
(Biomedical Center (BMC). Department of Physiological Chemistry. Faculty of Medicine. LMU Munich)
| Fecha: |
2023 |
| Resumen: |
All vertebrate genomes encode for three large histone H2A variants that have an additional metabolite-binding globular macrodomain module, macroH2A. MacroH2A variants impact heterochromatin organization and transcription regulation and establish a barrier for cellular reprogramming. However, the mechanisms of how macroH2A is incorporated into chromatin and the identity of any chaperones required for histone deposition remain elusive. Here, we develop a split-GFP-based assay for chromatin incorporation and use it to conduct a genome-wide mutagenesis screen in haploid human cells to identify proteins that regulate macroH2A dynamics. We show that the histone chaperone ANP32B is a regulator of macroH2A deposition. ANP32B associates with macroH2A in cells and in vitro binds to histones with low nanomolar affinity. In vitro nucleosome assembly assays show that ANP32B stimulates deposition of macroH2A-H2B and not of H2A-H2B onto tetrasomes. In cells, depletion of ANP32B strongly affects global macroH2A chromatin incorporation, revealing ANP32B as a macroH2A histone chaperone. |
| Ayudas: |
European Commission 804182
European Commission 851564
Agencia Estatal de Investigación PID2021-126907NB-I00
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| Nota: |
We would like to thank Carla Margulies, all members of the Ladurner and Mattiroli labs, and the Department of Physiological Chemistry for productive discussions. Thanks to Jan Dreyer, Inge Rondeel, and Rosanne van Hooijdonk for assistance with experiments. We thank the Bioimaging, Biophysics, and Flow Cytometry Core Facilities of the LMU Biomedical Center for training and use of their resources. We acknowledge the Protein Analytics Unit at the Biomedical Center, Ludwig-Maximilians University Munich (DFG, RI-00089), for providing services and assistance with data analysis. We thank the Histone Source at Colorado State University for the purification of human histones. pCAGGS-ANP32B was a kind gift from Wendy Barclay and pET29a-YS14 was a kind gift from Jung-Hyun Min (Addgene plasmid # 66890). pQCXIP-GFP1-10 was a gift from Yutaka Hata (Addgene plasmid # 68715) and pRK-flag-GFP11 from Yihong Ye (Addgene plasmid # 78590). This work was supported by funding from the Dutch Research Council (VI.Veni.212.052 to I.K.M.), the European Commission (ERC StG 851564 to F.M.; ERC StG 804182 to L.T.J.), the DFG (German Research Foundation) through Project-ID 213249687 - SFB 1064 and Project-ID 325871075 - SFB 1309, as well as LMU (to A.G.L.) and the national grant PID2021-126907NB-I00 from FEDER/Ministerio de Ciencia e Innovación (MCIN) - Agencia Estatal de Investigación and the Fundació La Marató de TV3 257/C/2019 (to M.B.). |
| Nota: |
We would like to thank Carla Margulies, all members of the Ladurner and Mattiroli labs, and the Department of Physiological Chemistry for productive discussions. Thanks to Jan Dreyer, Inge Rondeel, and Rosanne van Hooijdonk for assistance with experiments. We thank the Bioimaging, Biophysics, and Flow Cytometry Core Facilities of the LMU Biomedical Center for training and use of their resources. We acknowledge the Protein Analytics Unit at the Biomedical Center, Ludwig-Maximilians University Munich (DFG, RI-00089), for providing services and assistance with data analysis. We thank the Histone Source at Colorado State University for the purification of human histones. pCAGGS-ANP32B was a kind gift from Wendy Barclay and pET29a-YS14 was a kind gift from Jung-Hyun Min (Addgene plasmid # 66890). pQCXIP-GFP1-10 was a gift from Yutaka Hata (Addgene plasmid # 68715) and pRK-flag-GFP11 from Yihong Ye (Addgene plasmid # 78590). This work was supported by funding from the Dutch Research Council (VI.Veni.212.052 to I.K.M.), the European Commission (ERC StG 851564 to F.M.; ERC StG 804182 to L.T.J.), the DFG (German Research Foundation) through Project-ID 213249687 - SFB 1064 and Project-ID 325871075 - SFB 1309, as well as LMU (to A.G.L.) and the national grant PID2021-126907NB-I00 from FEDER/Ministerio de Ciencia e Innovación (MCIN) - Agencia Estatal de Investigación and the Fundació La Marató de TV3 257/C/2019 (to M.B.). Conceptualization: I.K.M. and A.G.L.; methodology: I.K.M. C.R. F.M. E.F. and L.T.J.; investigation: I.K.M. E.F. D.C. and C.K.; resources: C.R.; writing - original draft: I.K.M.; writing - review and editing: F.M. and A.G.L.; supervision: L.T.J. F.M. M.B. and A.G.L.; funding acquisition: I.K.M. L.T.J. M.B. F.M. and A.G.L. A.G.L. is a founder, CSO, and managing director of Eisbach Bio GmbH, a biotechnology company in oncology and virology. We support inclusive, diverse, and equitable conduct of research. |
| Derechos: |
Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.  |
| Lengua: |
Anglès |
| Documento: |
Article ; recerca ; Versió publicada |
| Materia: |
MacroH2A ;
Histone chaperone ;
ANP32B ;
Chromatin ;
Histone |
| Publicado en: |
Cell reports, Vol. 42 Núm. 10 (31 2023) , p. 113300, ISSN 2211-1247 |
visitante ::
identificación
(Institut Germans Trias i Pujol. Institut de Recerca contra la Leucèmia Josep Carreras)
