SETD3 is an Actin Histidine Methyltransferase that Prevents Primary Dystocia

For over fifty years, the methylation of mammalian actin at histidine 73 (actin-H73me) has been known to exist(1). Beyond mammals, we find that actin-H73me is conserved in several additional model animal and plant organisms. Despite the pervasiveness of H73me, its function is enigmatic, and the enzyme generating this modification is unknown. Here, we identify SETD3 (SET domain protein 3) as the physiologic actin histidine 73 methyltransferase. Structural studies reveal that an extensive network of interactions clamps the actin peptide on the SETD3 surface to properly orient H73 within the catalytic pocket and facilitate methyl transfer. H73me reduces the nucleotide exchange rate on actin monomers and modestly accelerates actin filament assembly. Mice lacking SETD3 show complete loss of actin-H73me in multiple tissues and quantitative proteomics singles out actin-H73 as the principal physiologic SETD3 substrate. SETD3 deficient female mice have severely decreased litter sizes due to primary maternal dystocia that is refractory to ecbolic induction agents. Further, depletion of SETD3 impairs signal-induced contraction in primary human uterine smooth muscle cells. Together, our results identify the first mammalian protein histidine methyltransferase and uncover a pivotal role for SETD3 and actin-H73me in the regulation of smooth muscle contractility. Our data also support the broader hypothesis where protein histidine methylation acts as a common regulatory mechanism.