A disordered N-terminal region of Ddx4 forms membraneless organelles in both cells and in vitro. These organelles are stabilized by electrostatic interactions and can be dissolved by changes in ionic strength, temperature, or arginine methylation. The study shows that phase separation of disordered proteins is a general mechanism for forming regulated, membraneless organelles. The Ddx4 N-terminal region, which is intrinsically disordered, forms droplets that concentrate single-stranded DNA but exclude double-stranded DNA, suggesting a role in nucleic acid localization. The droplets are dynamic, liquid-like structures that respond rapidly to environmental changes. The formation of these organelles is driven by sequence determinants that are common in membraneless organelle proteins. The study also reveals that post-translational modifications, such as arginine methylation, can regulate droplet formation. The sequence features that enable droplet formation in Ddx4 are found in other proteins associated with membraneless organelles, indicating that these features are general. The findings suggest that phase separation of disordered proteins is a widespread mechanism for forming membraneless organelles, providing a dynamic and responsive strategy for cellular compartmentalization.A disordered N-terminal region of Ddx4 forms membraneless organelles in both cells and in vitro. These organelles are stabilized by electrostatic interactions and can be dissolved by changes in ionic strength, temperature, or arginine methylation. The study shows that phase separation of disordered proteins is a general mechanism for forming regulated, membraneless organelles. The Ddx4 N-terminal region, which is intrinsically disordered, forms droplets that concentrate single-stranded DNA but exclude double-stranded DNA, suggesting a role in nucleic acid localization. The droplets are dynamic, liquid-like structures that respond rapidly to environmental changes. The formation of these organelles is driven by sequence determinants that are common in membraneless organelle proteins. The study also reveals that post-translational modifications, such as arginine methylation, can regulate droplet formation. The sequence features that enable droplet formation in Ddx4 are found in other proteins associated with membraneless organelles, indicating that these features are general. The findings suggest that phase separation of disordered proteins is a widespread mechanism for forming membraneless organelles, providing a dynamic and responsive strategy for cellular compartmentalization.