H4S47 O-GlcNAcylation regulates the activation of mammalian replication origins

Here, using proteomics, molecular biology and sequencing, the authors demonstrate that OGT-dependent O-GlcNAcylation of H4S47 is important for replication-origin activation, presumably through regulating DDK-induced phosphorylation of the MCM complex. The transmission and maintenance of genetic information in eukaryotic cells relies on the faithful duplication of the entire genome. In each round of division, excessive replication origins are licensed, with only a fraction activated to give rise to bi-directional replication forks in the context of chromatin. However, it remains elusive how eukaryotic replication origins are selectively activated. Here we demonstrate that O-GlcNAc transferase (OGT) enhances replication initiation by catalyzing H4S47 O-GlcNAcylation. Mutation of H4S47 impairs DBF4-dependent protein kinase (DDK) recruitment on chromatin, causing reduced phosphorylation of the replicative helicase mini-chromosome maintenance (MCM) complex and compromised DNA unwinding. Our short nascent-strand sequencing results further confirm the importance of H4S47 O-GlcNAcylation in origin activation. We propose that H4S47 O-GlcNAcylation directs origin activation through facilitating MCM phosphorylation, and this may shed light on the control of replication efficiency by chromatin environment.

Automated summary

Using proteomics, molecular biology and sequencing, the authors demonstrate the importance of OGT-dependent O-GlcNAcylation of H4S47 in the activation of replication origins. This occurs through regulation of DDK-induced phosphorylation of the MCM complex. The findings suggest that H4S47 O-GlcNAcylation enhances replication initiation by facilitating MCM phosphorylation, providing insight into the control of replication efficiency by the chromatin environment.