Biology subjects

Poppel, A. J.

Publications and source records attributed to Poppel, A. J..

3 recordsLinked to original sources

bioRxiv2026

Histone succinylation directly inhibits Jumonji domain demethylases and stabilizes repressive chromatin states

Herein we uncover a relationship between histone succinylation and Jumonji (JmjC) domain-containing histone demethylases. We used quantitative proteomics and peptide pull-down assays to identify JmjC demethylases as candidate interactors with succinylated histone peptides. Succinyl-lysine peptides bind and inhibit the catalytic activity of JmjC demethylases in a dose-dependent manner. This includes KDM4D and KDM6B, which are responsible for the removal of the silencing marks H3K9me2/3 and H3K27me2/3. Supraphysiological sodium succinate treatment of HepG2/C3A cells increased the relative abundance of histone succinylation, H3K9me2/3, and H3K27me2/3. CUT&Tag and ChIP-mass spectrometry revealed the co-occurrence of succinylation with these repressive methylation marks, in addition to reduced transcriptional output. This work establishes a novel mechanistic link between metabolite abundance and chromatin regulation and suggests a role for histone succinylation in the maintenance of heterochromatin.

biochemistry↗

bioRxiv2025

Crossover formation and coordinated assembly of synaptonemal complex relies on a direct interaction between Zip1 and Zip3

Several proteins collaborate to promote the crossover recombination events critical for accurate chromosome segregation during meiosis. How these "ZMM" factors (Zip2, Zip3, Zip4, Spo16, Mer3 and MutS{gamma}) collaboratively function remains incompletely understood. We previously reported that Zip3s abundance and activity rely on the synaptonemal complex (SC) component Zip1, and specifically on Zip1s N-terminal residues associated with crossovers and coupling SC assembly to the crossover pathway. Here, we demonstrate that Zip3 co-immunoprecipitates Zip1 from meiotic cells independent of recombination initiation and other ZMMs, and that Zip3s interaction with Zip1 relies on Zip1s N terminal residues. Co-expression and pull-down experiments in bacterial cells demonstrate that Zip1 and Zip3 interact directly. Experiments to identify Zip3 regions required for the Zip1 interaction unexpectedly revealed an incorrectly annotated translational start; we also determined that Zip3s N-terminal structured region is necessary and sufficient for the interaction, and a predicted coil downstream of Zip3s RING domain is essential for specific activities attributed to Zip1s N-terminal tip such as proximity labeling of Zip3 by Zip2 and the coupling of crossover recombination to SC assembly. Finally, we discovered that interaction with Zip1 protects Zip3 not only from proteasome-mediated degradation but also from post-translational modification when another ZMM is absent. We propose that direct interaction with Zip1s N terminus orients Zip3 within a nascent ZMM ensemble in a manner that facilitates crossover formation and the coupling of crossover intermediates to SC assembly, and furthermore ensures Zip3 remains both abundant and unmodified until all requisite ZMMs have joined the group.

genetics↗

bioRxiv2023

Proximity labeling reveals new insights into the relationships between meiotic recombination proteins in S. cerevisiae

Several protein ensembles facilitate MutS{gamma} crossover recombination and the associated process of synaptonemal complex (SC) assembly during meiosis, but the physical and functional relationships between the components involved remain obscure. We have employed proximity labeling as a phenotypic tool to discern functional relationships between meiotic recombination and SC proteins in S. cerevisiae, and to gain deeper insight into molecular deficits of crossover-defective mutants. We find that recombination initiation (Spo11) and the Mer3 helicase are dispensable for proximity labeling of the Zip3 E3 ligase by components of the ZZS ensemble (Zip2, Zip4 and Spo16) but are required for proximity labeling of Zip3 by Msh4, consistent with the possibility that MutS{gamma} joins Zip3 only after a specific recombination intermediate has been generated. Proximity labeling analysis of crossover-defective zip1 mutants suggests a key shared defect is a failure to assemble an early recombination ensemble where ZZS can properly engage Zip3. We furthermore discovered that Zip3s abundance within the meiotic cell is uniquely dependent on the presence of Zip1, and that the post-translational modification of Zip3 is promoted by most MutS{gamma} pathway proteins but countered by Zip1. Based on this and additional data, we propose a model whereby Zip1 stabilizes a functional, unmodified form of Zip3 until intermediate steps in recombination are complete. We also find that SC structural protein Ecm11 is proximity labeled by ZZS complex proteins in a Zip4-dependent manner, but by Zip3 and Msh4, at least in part, via a distinct pathway. Finally, streptavidin pulldowns followed by mass spectrometry on eleven different proximity labeling strains uncovers shared proximity targets of MutS{gamma}-associated proteins, some with known meiotic functions and others not yet implicated in a meiotic activity, highlighting the potential power of proximity labeling as a discovery tool.

genetics↗