Bipolar switching by HCN voltage sensor underlies hyperpolarization activation

Hyperpolarization-activated, cyclic nucleotide-gated (HCN) channels show an inverted voltage response compared with virtually all other voltage-gated channels, opening on hyperpolarization rather than depolarization. Although the structure of the HCN1 channel was recently solved, the structural element(s) responsible for the inverted gating polarity of HCN is not known. Here, we use a hierarchical approach, by first characterizing the functional contribution of each structural element to channel gating, and then identifying the critical interactions between these elements. Our studies reveal that the HCN voltage sensor can gate the same pore open on both depolarization and hyperpolarization, thereby acting as a bipolar switch. Elements in the pore domain shut off the depolarization-activation pathway in wild-type channels.