In addition to Natchez and Nashville, larger cities along the route include Jackson and Tupelo, Mississippi, and Florence, Alabama.

Its design harkens back to the way the original interweaving trails aligned as an ancient salt lick-to-grazing pasture migratory route of the American bison and other game that moved between grazing the pastures of central and western Mississippi and the salt and other mineral surface deposits of the Cumberland Plateau.

The route generally traverses the tops of the low hills and ridges of the watershed divides from northeast to southwest.

Native Americans, following the "traces" of bison and other game, further improved this walking trail for foot-borne commerce between major villages located in central Mississippi and middle Tennessee.

Also avoided was the danger to a herd (or groups of human travelers) of being caught en masse at the bottom of a hollow or valley if attacked by predators.

In the early post-American Revolutionary War period of America's (south) westward expansion, the trace was the return route for American flat-boat commerce between the territories of the upper and lower Ohio, Tennessee, and Cumberland River valleys.

The Americans constructed flat boats, loaded their commerce therein, and drifted upon those rivers, one-way south-southwestward to New Orleans, Louisiana.

Improved communications (steam boats, stagecoach lines, and railroads) and the development of ports along the rivers named above (e.g., Natchez; Memphis, Tennessee; Paducah, Kentucky; Nashville, Tennessee; and Louisville, Kentucky) made the route obsolete as a means of passenger and freight commerce.

Construction of the parkway was begun by the federal government in the 1930s, one of the many projects of the Civilian Conservation Corps during the Great Depression.

In 2013, a new law required the National Park Service (NPS) to convey about 67 acres of property in the Natchez Trace Parkway to the State of Mississippi.

The two pieces of land in question originally belonged to Mississippi and were donated to the NPS when it was trying to determine where to end the Natchez Trace Parkway.

The mound was built by depositing earth along the sides of a natural hill, thus reshaping it and creating an enormous artificial plateau.