The bus, travelling from Boisar in the district to Swargate bus stand in Pune, collided with a dumper on Mumbai-Ahmedbad highway at around 3.30 pm.
"The accident took place near Maljipada village between Vasai and Ghodbunder in Palghar district, in which the driver and conductor of the Maharashtra State Road Transport Corporation (MSRTC) bus died on the spot," Vasai tahsildar Gajendra Patole said.
Some of the occupants of the bus were also injured in the accident, the official said.