A motif in HSP90 and P23 that links molecular chaperones to efficient estrogen receptor α methylation by the lysine methyltransferase SMYD2

Heat shock protein 90 (HSP90) is a molecular chaperone that supervises folding of cellular signaling proteins such as steroid receptors and many protein kinases. HSP90 relies on ATP hydrolysis for powering a conformational circuit that helps fold the client protein. To that end, HSP90 binds to co-chaperone proteins that regulate ATP hydrolysis rate or interaction with client proteins. Co-chaperones such as P23, cell division cycle 37 (CDC37), or activator of HSP90 ATPase activity 1 (AHA1) interact with the N-terminal or middle domain of HSP90, whereas others, such as HSP70/HSP90-organizing protein (HOP), use tetratricopeptide repeat (TPR) domains to bind the EEVD motif at the very C-terminal end of HSP90. Recently, the lysine methyltransferase SET and MYND domain-containing 2 (SMYD2) has been proposed as an HSP90-binding partner, and interaction analyses indicate that SMYD2 binding to HSP90 is independent of the EEVD motif. Using the amplified luminescence proximity homogeneous assay (Alpha) technique, I identified a new (M/I/L/V)PXL motif at the C termini of HSP90 and P23 that mediates an interaction with SMYD2, and synthetic peptides harboring this motif dissociated this complex. Of note, the HSP90- and P23-dependent client estrogen receptor (ER), was a major methylation target of SMYD2. In a reconstituted system in bacteria, I analyzed HSP90/P23-associated, SMYD2-mediated ER methylation and found that when SMYD2 binds to the molecular chaperones, it considerably increases methylation of Lys-266 in ER. Because methylation represses ER activity, the observed complex formation between SMYD2 and HSP90/P23 may contribute to ER regulation.