The town's branch of Homebase will shut its doors for the last time in September.
A spokesman for the chain said: "We can confirm that the lease has expired at our Doncaster store and will close at the end of September.
"We are continuing to support team members by helping them find alternative employment and customers will still be able to shop at the nearby Sheffield or Selby stores."
The firm was sold by its owner Home Retail Group to Australian retail giant Westfarmers for £340m in 2016.
The firm also owns the Australian DIY chain Bunnings and all 265 of the UK's Homebase sites are being rebranded under the Bunnings banner over the next three to five years.
Prior to being known as Homebase, the store in Milethorn Lane was known as Texas Homecare.