AC6 regulates the microtubule-depolymerizing kinesin KIF19A to control ciliary length in mammals.

Motile cilia are hairlike structures that line the respiratory and reproductive tracts and the middle ear and generate fluid flow in these organs via synchronized beating. Cilium growth is a highly regulated process that is assumed to be important for flow generation. Recently, Kif19a, a kinesin residing at the cilia tip, was identified to be essential for ciliary length control through its microtubule depolymerization function. However, there is a lack of information on the nature of proteins and the integrated signaling mechanism regulating growth of motile cilia. Here, we report that adenylate cyclase 6 (AC6), a highly abundant AC isoform in airway epithelial cells, inhibits degradation of Kif19a by inhibiting autophagy, a cellular recycling mechanism for damaged proteins and organelles. Using epithelium-specific knockout mice of AC6, we demonstrated that AC6 knockout airway epithelial cells have longer cilia compared with the WT cells because of decreased Kif19a protein levels in the cilia. We demonstrated in vitro that AC6 inhibits AMP-activated kinase (AMPK), an important modulator of cellular energy-conserving mechanisms, and uncouples its binding with ciliary kinesin Kif19a. In the absence of AC6, activation of AMPK mobilizes Kif19a into autophagosomes for degradation in airway epithelial cells. Lower Kif19a levels upon pharmacological activation of AMPK in airway epithelial cells correlated with elongated cilia and vice versa. In all, the AC6-AMPK pathway, which is tunable to cellular cues, could potentially serve as one of the crucial ciliary growth checkpoints and could be channeled to develop therapeutic interventions for cilia-associated disorders.

Intramolecular ß-Alkenylation of Cyclohexanones via Pd-Catalyzed Desaturation-Mediated C(sp3)-H/Alkyne Coupling.

Site-selective C-C bond formation through the direct coupling of C(sp3)-H bonds with unsaturated hydrocarbons represents an atom-economical and redox-neutral way to functionalize chemically inert positions, such as those ß to a carbonyl group. While most existing ß-functionalization methods utilize a directing group (DG) strategy, here we report a Pd-catalyzed intramolecular ß-alkenylation of ketones using alkynes as the coupling partner without the aid of DGs. Mediated by a ketone desaturation process, the reaction is redox-neutral and avoids using strong acids or bases. The resulting cis-5,6-fused bicycles can be diversely derivatized with excellent selectivity. Mechanistic studies imply an unusual "hydride-transfer" chain-like pathway, which involves the cyclometalation of an enyne intermediate and protonation of the resulting Pd enolate followed by an intermolecular hydride transfer through the desaturation of another substrate.