DS0405 -

Integrative molecular, functional and structural analysis of histone variant H2A.Z – EpiVarZ

Submission summary

Histone variants are major epigenetic regulators of transcription, DNA replication, repair and recombination. Their deposition and eviction from specific sites on chromatin are highly regulated processes mediated by specific chaperones often part of larger chromatin remodelling/modification complexes. How these molecular machines selectively recognise histone variants and the mechanism and regulation of deposition and eviction from chromatin are critical questions that must be addressed.
H2A.Z1 and H2A.Z2 are two highly related variants of canonical H2A. The authors of this proposal have made seminal discoveries on how histone chaperones YL1 and ANP32E selectively recognise H2A.Z1 to deposit and evict it from the nucleosome. Building on these studies, we aim here to address how these processes are regulated, to identify additional chaperones and to define the functions of H2A.Z1 and H2A.Z2 in mitosis, development, tissue homeostasis and tumourigenesis.
Approaches using genetically modified MEFs in which the genes encoding these variants carry a tag will serve to identify specific histone chaperones for H2A.Z1 and H2A.Z2. This approach will also be used to identify chaperones and complexes involved in deposition/eviction of these variants specifically during mitosis and to define the localisation of each variant on the genome throughout the cell cycle. The components of novel complexes, in particular the specific chaperones, will be identified, their structure solved by X-ray crystallography and the basis of specific recognition of H2A.Z1 and H2A.Z2 defined. We will define how the YL1 and ANP32E chaperones interact with the SRCAP and P400 complexes, identifying their direct interaction partners and defining whether and how integration modifies their chaperone activity. We will use mice bearing conditional floxed alleles of the H2afz1 and H2afz2 genes to inactivate the corresponding proteins in adult hepatocytes addressing their roles in tissue homeostasis and in a mouse model of melanoma defining their roles in tumourigenesis. We will also generate MEFs and embryonic stem cells where the H2afz1 and/or H2afz2 can be inactivated to address their roles in mitosis, viability, pluripotency and differentiation.
This project presents functional, structural and genetic approaches to address critical questions on the H2A.Z variants from the atomic structure of their chaperones, the biochemistry of their deposition and eviction from the nucleosome to their functions in mouse tissues and tumourigenesis. We believe that we have formed a unique consortium to address these issues in an integrative and comprehensive manner leading to a new and unprecedented level of understanding of these important histone variants.

Project coordination

Ali HAMICHE (Institut de Génétique et de Biologie Moléculaire et Cellulaire)

The author of this summary is the project coordinator, who is responsible for the content of this summary. The ANR declines any responsibility as for its contents.

Partner

IGBMC Institut de Génétique et de Biologie Moléculaire et Cellulaire
IGBMC Institut de Génétique et de Biologie Moléculaire et Cellulaire
IAB - Equipe DIMITROV Institut Albert Bonniot - CR Inserm U1209/ CNRS UMR 5309 / UGA
IGBMC Institut de Génétique et de Biologie Moléculaire et Cellulaire

Help of the ANR 628,617 euros
Beginning and duration of the scientific project: September 2016 - 36 Months

Useful links

Explorez notre base de projets financés

 

 

ANR makes available its datasets on funded projects, click here to find more.

Sign up for the latest news:
Subscribe to our newsletter