Histone H3 Ser57 and Thr58 phosphorylation in the brain of 5XFAD mice

• We used a mouse model of rapid amyloid deposition (5XFAD) to examine histone phosphorylation in the brain.
• We measured phosphorylation of histone H3 Ser-57 and Thr-58 using multiple reaction monitoring (MRM) mass spectrometry.
• Decrease in phosphorylation could increase DNA–histone interactions and condense chromatin, reducing gene expression.
• MRM was also used to perform absolute quantification of amyloid precursor protein and amyloid-β in mouse brain.

Alzheimer’s disease has been shown to have a global reduction in gene expression, called an epigenetic blockade, which may be regulated by histone post-translational modifications. Histone H3 has been shown to be highly regulated by phosphorylation. We, therefore, chose H3 for investigation of phosphorylation of the core sites serine-57 (S57) and threonine-58 (T58). Hemispheres of brains from a mouse model of rapid amyloid deposition (5XFAD) were used for measurement of S57 and T58 phosphorylation. Multiple reaction monitoring (MRM) was used to measure the level of phosphorylation, which was normalized to a non-modified “housekeeping” peptide of H3. S57 phosphorylation was decreased by 40%, T58 phosphorylation was decreased by 45%, and doubly phosphorylated S57pT58p was decreased by 30% in 5XFAD brain in comparison to C57BL/6J age- and sex-matched wild type controls. Amyloid-β (Aβ) and amyloid precursor protein were also measured to confirm that 5XFAD mice produced high levels of Aβ. Decreased phosphorylation of these sites in close proximity to DNA may lead to stabilization of DNA–histone interactions and a condensed chromatin state, consistent with the epigenetic blockade associated with AD. Our findings of H3 sites S57 and T58 exhibiting lower levels of phosphorylation in 5XFAD model compared to wild type control implicate these sites in the epigenetic blockade in neurodegeneration pathology.

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