Condensin I^DChas a Functional ATPase That is Required for X-Chromosome Dosage Compensation inC. elegans

Dosage compensation (DC) inC. elegansutilizes a condensin complex that resembles mitotic condensins, but differs by one subunit, DPY-27. DPY-27 replaces SMC-4, one of the Structural Maintenance of Chromosome (SMC) proteins that is responsible for hydrolyzing ATP, required for condensation of DNA and other mitotic condensin functions. To understand if the ATPase function is required in DC, we first demonstrated that DPY-27 is capable of hydrolyzing ATPin vitro. Then, we used CRISPR/Cas9-mediated genome editing to generate an ATPase mutation indpy-27and demonstrated that this mutation results in a loss of DC. Specifically, we found that without ATPase function, DPY-27 containing condensin I^DChas reduced capacity to bind DNA, condense the X chromosomes, and facilitate H4K20me1 enrichment on the X-chromosomes. Our results suggest that condensin I^DC, like mitotic condensins, uses ATP hydrolysis to perform its functions, makingC. elegansDC a model for how activities attributed to mitotic condensins can be used to regulate gene expression.