Biology subjects

Katch, B. J.

Publications and source records attributed to Katch, B. J..

3 recordsLinked to original sources

bioRxiv2026

KAT6 and KAT7 actively maintain mitotic chromatin composition and acetylation to preserve developmental competence

Mitotic bookmarking enables transmission of regulatory information across cell division, yet how active chromatin states are molecularly preserved during mitosis and how they contribute to postmitotic cell fate decisions remain unresolved. Here, we demonstrate that the acetyltransferases KAT6 and KAT7 remain chromatin-bound and catalytically active throughout mitosis in mouse and human pluripotent stem cells. Acute KAT6/7 depletion disrupts mitotic histone acetylation and the retention of multiple chromatin regulators with minimal effects on chromatin accessibility. Mechanistically, KAT6/7 bookmark both self renewal and developmental gene loci, where they are required for post-mitotic transcriptional reactivation and priming of lineage-specific programs. Consistently, genes impaired during postmitotic reactivation also fail to activate during neuronal differentiation upon continuous KAT6/7 perturbation, compromising the establishment of the neuronal program and identity. Altogether, our findings identify KAT6 and KAT7 as a cooperative mitotic chromatin maintenance system that couples active histone acetylation to preservation of chromatin composition, transcriptional memory, and developmental competence across cell division, extending the bookmarking function beyond transcriptional memory. These findings also provide a new framework for understanding KAT6/7 function in neurodevelopmental disorders and cancer.

molecular biology↗

bioRxiv2025

A non-catalytic role for RFC in PCNA-mediated processive DNA synthesis

The ring-shaped sliding clamp PCNA enables DNA polymerases to perform processive DNA synthesis during replication and repair. The loading of PCNA onto DNA is catalyzed by the ATPase clamp loader RFC. Using a single-molecule platform to visualize the dynamic interplay between PCNA and RFC on DNA, we unexpectedly discovered that RFC continues to associate with PCNA after loading, contrary to the conventional view. Functionally, this clamp-loader/clamp complex is required for processive DNA synthesis by polymerase {delta} (Pol{delta}), as the PCNA-Pol{delta} assembly is inherently unstable. This architectural role of RFC is dependent on the BRCT domain of Rfc1, and mutation of its DNA-binding residues causes sensitivity to DNA damage in vivo. We further showed the FEN1 flap endonuclease can also stabilize the PCNA-Pol{delta} interaction and mediate robust synthesis. Overall, our work revealed that, beyond their canonical enzymatic functions, PCNA-binding proteins harbor non-catalytic functions essential for DNA replication and genome maintenance.

biophysics↗

bioRxiv2023

Synergistic Binding of the Halide and Cationic Prime Substrate of the L-Lysine 4-Chlorinase, BesD, in Both Ferrous and Ferryl States

An aliphatic halogenase requires four substrates: 2-oxoglutarate (2OG), halide (Cl- or Br-), the halogenation target ("prime substrate"), and dioxygen. In well-studied cases, the three non-gaseous substrates must bind to activate the enzymes Fe(II) cofactor for efficient capture of O2. Halide, 2OG, and (lastly) O2 all coordinate directly to the cofactor to initiate its conversion to a cis-halo-oxo-iron(IV) (haloferryl) complex, which abstracts hydrogen (H*) from the non-coordinating prime substrate to enable radicaloid carbon-halogen coupling. We dissected the kinetic pathway and thermodynamic linkage in binding of the first three substrates of the O_SCPLOWLC_SCPLOW-lysine 4-chlorinase, BesD. After 2OG adds, subsequent coordination of the halide to the cofactor and binding of cationic O_SCPLOWLC_SCPLOW-Lys near the cofactor are associated with strong heterotropic cooperativity. Progression to the haloferryl intermediate upon addition of O2 does not trap the substrates in the active site and, in fact, markedly diminishes cooperativity between halide and O_SCPLOWLC_SCPLOW-Lys. The surprising lability of the BesD*[Fe(IV)=O]*Cl*succinate*O_SCPLOWLC_SCPLOW-Lys complex engenders pathways for decay of the haloferryl intermediate that do not result in O_SCPLOWLC_SCPLOW-Lys chlorination, especially at low chloride concentrations; one identified pathway involves oxidation of glycerol. The mechanistic data imply that (i) BesD may have evolved from a hydroxylase ancestor either relatively recently or under weak selective pressure for efficient chlorination and (ii) that acquisition of its activity may have involved the emergence of linkage between O_SCPLOWLC_SCPLOW-Lys binding and chloride coordination following loss of the anionic protein-carboxylate iron ligand present in extant hydroxylases.

biochemistry↗