Peripheral sequestration of huntingtin delays neuronal death and depends on N-terminal ubiquitination

Abstract

Huntington’s disease (HD) is caused by a glutamine repeat expansion in the protein huntingtin. The mutated protein (mHtt) forms aggregates whose impacts on neuronal survival are still debated. Using weeks-long, continual imaging of individual cortical neurons, we find that mHtt is gradually sequestrated into peripheral, mainly axonal aggregates, concomitant with dramatic reductions in cytosolic mHtt levels and enhanced neuronal survival. in-situ pulse-chase imaging reveals that aggregates continually gain and lose mHtt, in line with these acting as mHtt sinks at equilibrium with cytosolic pools. Preventing ubiquitination at two N-terminal lysines observed only in HD animal models suppresses peripheral aggregate formation and reductions in cytosolic mHtt, promotes nuclear aggregate formation, stabilizes aggregates and leads to pervasive neuronal death. These findings demonstrate the capacity of aggregates formed at peripheral locations to sequester away cytosolic, presumably toxic mHtt forms and support a crucial role for N-terminal ubiquitination in promoting these processes and delaying neuronal death.