On the night of November 4th going into the 5th, 1960, Horton and two other band members, Tommy Tomlinson and Tillman Franks, were traveling from the Skyline Club in Austin, Texas to Shreveport when they collided with an oncoming truck on a bridge near Milano in Milam County, Texas. Horton died en route to the hospital, and Tomlinson was seriously injured; his leg was later amputated. Franks suffered head injuries, and James Davis, the driver of the truck, sustained a broken ankle and other minor injuries.

The funeral was held in Shreveport on November 8, 1960, officiated by Franks' younger brother William Derrel "Billy" Franks, a Church of God minister. Johnny Cash performed one of the readings, choosing Chapter 20 from the Gospel of John (Resurrection of Jesus).

When Johnny Cash, a good friend of Horton's, learned about the accident, he said, "I locked myself in one of the hotel's barrooms and cried." Cash dedicated his rendition of "When It's Springtime in Alaska (It's Forty Below)" to Horton on his album Personal File: "Johnny Horton was a good old friend of mine."