Phosphorylation of histone H3 Ser10 establishes a hierarchy for subsequent intramolecular modification events

Abstract

Phosphorylation of Ser10 of histone H3 regulates chromosome condensation and transcriptional activity. Using time-resolved, high-resolution NMR spectroscopy, we demonstrate that histone H3 Ser10 phosphorylation inhibits checkpoint kinase 1 (Chk1)- and protein kinase C (PKC)-mediated modification of Thr11 and Thr6, the respective primary substrate sites of these kinases. On unmodified H3, both enzymes also target Ser10 and thereby establish autoinhibitory feedback states on individual H3 tails. Whereas phosphorylated Ser10 does not affect acetylation of Lys14 by Gcn5, phosphorylated Thr11 impedes acetylation. Our observations reveal mechanistic hierarchies of H3 phosphorylation and acetylation events and provide a framework for intramolecular modification cross-talk within the N terminus of histone H3.

The Bromodomain of Gcn5 Regulates Site Specificity of Lysine Acetylation on Histone H3

“…Phosphorylation of threonine 11 on histone H3 reduces the ability of Gcn5 to acetylate H3K14 (80). NuA3, a yeast acetyltransferase complex containing the HAT Sas3, binds trimethylated H3K4 using the PHD finger of the Yng1 subunit, which stimulates H3K14 acetylation on the same H3 polypeptide (8,81).…”

Histone phosphorylation

“…recent in vitro findings using time-resolved NMr spectroscopy suggest a slightly modified picture in which H3S10ph acts as master switch for subsequent intramolecular modifications, and inhibits T6 and T11 phosphorylation while the reverse does not apply. 139 This study also questioned the role of T11ph in the stimulation of K14ac.…”

Differentially Isotope‐Labeled Nucleosomes To Study Asymmetric Histone Modification Crosstalk by Time‐Resolved NMR Spectroscopy

“…As before, kinetic analyses with these nucleosomes showed faster acetylation of the S10 prephosphorylated H3 copy ( Figure S6), thus confirming the stimulatory role of S10ph on Gcn5 activity in cis compared to trans. This effect is nucleosome-specific since Gcn5 processes both unmodified and H3S10ph tail peptides similarly [5] and is directly linked to the influence that charge-modulating PTMs have on electrostatic H3 tail/DNA interactions. Particularly, H3S10ph disrupts the aforementioned transient contacts, hence promotes K14ac by rendering H3 tail more accessible to Gcn5.…”

“…[4] Historically, and because of the inherent symmetry of the nucleosomal architecture, histone PTMs were thought to exclusively occur in a symmetrical fashion, with both copies of each of the core histones modified in exactly the same manner. This notion was recently challenged by two studies demonstrating the mechanistic basis for establishing asymmetric histone H3 phosphorylations in vitro by certain kinases, [5] and the existence of asymmetrically methylated nucleosomes in embryonic stem cells, fibroblasts and cancer cells. [6] As such, asymmetrically modified nucleosomes represent a novel concept in chromatin biology and their existence raises important biological questions with regard to their establishment and crosstalk in cis, i.e.…”