Neuronal SETD2 activity links microtubule methylation to an anxiety-like phenotype in mice

Brain. 2021 May 20;awab200. doi: 10.1093/brain/awab200. Online ahead of print.


Gene discovery efforts in autism spectrum disorder have identified heterozygous defects in chromatin remodeler genes, the" readers, writers and erasers" of methyl marks on chromatin, as major contributors to this disease. Despite this advance, a convergent etiology between these defects and aberrant chromatin architecture or gene expression has remained elusive. Recently, data have begun to emerge that chromatin remodelers also function directly on the cytoskeleton. Strongly associated with autism spectrum disorder, the SETD2 histone methyltransferase for example, has now been shown to directly methylate microtubules of the mitotic spindle. However, whether microtubule methylation occurs in post-mitotic cells, for example on the neuronal cytoskeleton, is not known. We found the SETD2 α-tubulin lysine 40 trimethyl mark occurs on microtubules in the brain and in primary neurons in culture, and that the SETD2 C-terminal SRI domain is required for binding and methylation of α-tubulin. A CRISPR knock-in of a pathogenic SRI domain mutation (Setd2SRI) that disables microtubule methylation revealed at least one wild-type allele was required in mice for survival, and while viable, heterozygous Setd2SRI/wt mice exhibited an anxiety-like phenotype. Finally, whereas RNA-seq and ChIP-seq showed no concomitant changes in chromatin methylation or gene expression in Setd2SRI/wt mice, primary neurons exhibited structural deficits in axon length and dendritic arborization. These data provide the first demonstration microtubules of neurons are methylated, and reveals a heterozygous chromatin remodeler defect that specifically disables microtubule methylation is sufficient to drive an autism-associated phenotype.

Keywords: SETD2; SRI domain; autism spectrum disorder; haploinsufficient; α-tubulin methylation.