This roadway was originally part of U.S. Route 65 until 1967, when a bypass was built around Western Grove. The main route was rerouted onto the bypass, with the downtown alignment becoming US 65 Business.
U.S. Route 65 Business in Springfield parallels US 65, which follows the Schoolcraft Freeway along the east edge of town, for its entire length. Route 65 Business begins at an interchange with US 60, the James River Freeway. For most of its length, the route follows Glenstone Avenue, a major retail corridor in Springfield. Glenstone Avenue is an expressway from US 60 to Battlefield Road. At the corner of Battlefield and Glenstone lies Battlefield Mall, the region's largest shopping mall.
Business 65 continues north as a four-lane arterial road. At Sunshine Street, it serves as the western terminus of Route D. Route 65 Business continues along Glenstone until the intersection with Chestnut Expressway, where it meets I-44 Business. Eastbound I-44 BUS turns north from Chestnut onto Glenstone at this intersection, while Business 65 turns east onto Chestnut. Thus, the two routes do not truly intersect; all motorists must make a turn to continue on the numbered route that they were previously on. Business 65 then proceeds eastward along Chestnut Expressway until it intersects with mainline US-65, where the business route terminates. Originally, Business 65 continued north on Glenstone to Interstate 44; the routing was shifted when Chestnut Expressway east of Glenstone was turned over to MoDOT by the city in exchange for Division Street (previously Missouri YY between US 65 and Glenstone).
U.S. Highway 65 Business starts at the junctions of Missouri Route 6 and US 65 and runs west a few hundred yards before turning north onto Oklahoma Avenue. It follows Oklahoma Avenue north to the junction of Oklahoma Avenue and 28th Street where it turns east and terminates at US 65.