Understanding the functional role of ZNF512B in NuRD-dependent and -independent chromatin processes

• Applicant: Professorin Dr. Sandra Brigitte Hake
• Subject Area: General Genetics and Functional Genome Biology
• Term: since 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 533588517

The conserved histone variant H2A.Z plays a crucial role in various DNA-based processes but the underlying mechanisms are not completely understood. Recently, we identified the zinc finger (ZNF) protein ZNF512B as H2A.Z associated protein. Here, we report ZNF512B as a novel nucleosome remodeling and deacetylase (NuRD) complex binding protein. We discovered a conserved amino acid sequence within ZNF512B that resembles a NuRD-interaction domain (NIM). By solving the crystal structure of this domain directly bound to the NuRD component RBBP4 and by applying several biochemical assays we demonstrate that this novel NIM sequence is both necessary and sufficient for NuRD binding. Transcriptome analyses and reporter assays identify ZNF512B as regulator (repressor) of gene expression, in both NuRD-dependent and -independent manners. Surprisingly, high levels of ZNF512B expression lead to nuclear protein aggregation and overlapping DNA/chromatin compaction that are independent of the interaction with the NuRD complex but depend on ZNF512B’s zinc fingers. In this DFG project, we plan to investigate whether ZNF512B's four typical C2H2 zinc fingers are involved in DNA binding and whether its four atypical C2HC zinc fingers mediate oligomerization, which can ultimately lead to protein and DNA/chromatin aggregation. Furthermore, using ZNF512B depleted mouse embryonic stem cells, we will determine both ZNF512B's DNA and chromatin binding specificities as well as its function as transcriptional regulator during neuronal differentiation. Our study will not only reveal the biological role of ZNF512B in chromatin compaction and differentiation processes, but also provide new insights into the impact of deregulated ZNF512B levels in diseases such as cancer and neurodegenerative diseases.

• DFG Programme: Research Grants