Local police say the deaths from cardiopulmonary arrest were confirmed by doctors at the scene.
The accident took place shortly before 5 AM on Sunday near a junction on the Kan-etsu expressway in Fujioka City, Gunma Prefecture.
Local fire and emergency authorities say at least nine people were seriously injured.
Pictures taken from an NHK helicopter show that the front part of the bus is badly damaged, and much of the vehicle is sticking into mid-air off the expressway.
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