U.S. Route 501 (US 501) is a part of the U.S. Highway System that runs from Myrtle Beach, South Carolina to Buena Vista, Virginia. In Virginia, the U.S. Highway runs 111.42 miles (179.31 km) from the North Carolina state line near Cluster Springs north to its northern terminus at US 60 in Buena Vista. US 501 is the primary highway between Lynchburg and both South Boston in Southside Virginia and Durham in North Carolina's Research Triangle. North of Lynchburg, the highway parallels the James River through the Blue Ridge Mountains to the southern end of the Shenandoah Valley.

US 501 enters Halifax County, Virginia south of Cluster Springs. The U.S. Highway is a two-lane road at the North Carolina state line but immediately expands to a four-lane divided highway named Huell Matthews Highway. US 501 parallels the Mayo River north to the hamlet of Mayo, where the highway meets the western end of SR 96 (Virgilina Road) and crosses the Hyco River. The U.S. Highway passes through Cluster Springs to the community of Riverdale just south of South Boston. US 501 becomes an undivided highway and intersects US 58 and US 360, which together head west as Philpott Road and east as Bill Tuck Highway. US 501 crosses the Dan River into the town of South Boston. Just north of the river, the highway has a grade crossing of Norfolk Southern Railway's Danville–Richmond rail line and splits into a one-way pair: Broad Street northbound and Main Street southbound. Both directions intersect SR 304 (Seymour Drive). When Main Street veers northeast as two-way SR 129, southbound US 501 continues on two-way Wilborn Avenue to the northern end of the one-way pair.

US 501 continues through South Boston on Wilborn Avenue, a five-lane road with center turn lane. The U.S. Highway becomes four-lane divided Halifax Road at Hamilton Boulevard. US 501 receives the northern end of SR 129 (Old Halifax Road) shortly before leaving the town of South Boston. The U.S. Highway enters the town of Halifax and becomes Main Street, a three-lane road with center turn lane, at its grade crossing of Norfolk Southern's Durham District. US 501 continues to the county courthouse, where the highway intersects the very short SR 349 (Edmunds Boulevard) and SR 360 (Mountain Road), which runs concurrently with the U.S. Highway through the northern part of the town. The two highways diverge: SR 360 heads northeast as Bethel Road and US 501 heads northwest as two-lane L.P. Bailey Memorial Highway. which crosses Banister Lake, an impoundment of the Banister River, as it leaves the town of Halifax.

US 501 passes through Volens and Acorn on its way to the hamlet of North Halifax, where the highway begins to run concurrently with SR 40 (Stage Coach Road). The highways cross the Roanoke River into the town of Brookneal in Campbell County and pass under Norfolk Southern's Altavista District rail line. The highways continue as Lusardi Drive to Lynchburg Avenue in the center of town; SR 40 turns southeast while US 501 turns north west onto the street just north of the highways' grade crossing of the Durham District rail line. US 501 leaves the town as Brookneal Highway, which parallels the rail line through the communities of Naruna and Gladys on its way to Rustburg, the county seat of Campbell County. The U.S. Highway intersects SR 24 (Colonial Highway) at the southern edge of the village. The highways have a short concurrency before SR 24 continues east as Village Highway and US 501 heads northwest as Campbell Highway. North of Rustburg, US 501 expands to a four-lane divided highway into the independent city of Lynchburg, where the highway meets US 29 and US 460 (Richmond Highway) at a partial cloverleaf interchange. US 501 Business and US 460 Business head straight on Campbell Avenue while US 501 joins the freeway heading southwest.

US 501 runs concurrently with US 29 and US 460 southwest along the northern slope of Candlers Mountain to a full Y interchange where US 501 heads northwest from the freeway. The U.S. Highway has an intersection with SR 128 (Mayflower Drive) and Candlers Mountain Road just east of the highway's crossing of Norfolk Southern's Danville District. SR 128 joins US 501 on Candlers Mountain Road between the intersection and the highway's cloverleaf interchange with US 29 Business (Lynchburg Expressway). US 501 exits onto the expressway while SR 128 continues west to its terminus. US 501 and US 29 Business head southwest along the freeway until the business route exits onto Wards Road to head toward Danville. US 501 curves northwest and has a cloverleaf interchange with US 460 Business (Timberlake Road), crosses Norfolk Southern's Blue Ridge District, and has a diamond interchange with Graves Mill Road. The freeway ends at US 221 (Lakeside Drive). US 501 heads north along Old Forest Road as a five-lane road with center turn lane before turning onto a two-lane segment of the Lynchburg Expressway. This segment has a partial cloverleaf interchange with Wiggington Road before ending at US 501 Business (Boonsboro Road).

US 501 heads northwest on two-lane Boonsboro Road, which becomes Lee Jackson Highway when it exits the city of Lynchburg into Bedford County. The U.S. Highway curvaceously crosses Fleming Mountain and descends into the James River gorge at Coleman Falls. US 501 follows the south bank of the river through Big Island, where the highway meets the northern end of SR 122 (Big Island Highway) and has an interchange with the Blue Ridge Parkway. North of Big Island, the U.S. Highway crosses the river and immediately intersects SR 130 (Elon Road) on the Amherst County side. The two highways have a curvaceous crossing of the spine of the Blue Ridge Mountains before descending to the town of Glasgow at the mouth of the Maury River. The two routes follow the river upstream a short distance before SR 130 (Wert Faulkner Highway) crosses the river into the town. US 501, now named Glasgow Highway, parallels the river and Norfolk Southern's Roanoke District rail line north to the city of Buena Vista, which the highway enters as Magnolia Avenue. South of the downtown area, US 501 veers onto Sycamore Street while US 501 Business continues along Magnolia Avenue. At 22nd Street, the U.S. Highway veers onto Beech Avenue, which receives the northern end of US 501 Business (Park Avenue) west of Southern Virginia University. Two blocks to the north, US 501 reaches its northern terminus at US 60 (29th Street).

U.S. Route 501 Business (US 501 Business) is a business route of US 501 in Lynchburg. The highway runs 9.41 miles (15.14 km) from US 29, US 460, and US 501 on the eastern edge of Lynchburg to US 501 on the western edge of Lynchburg. US 501 Business runs concurrently with US 460 Business on Campbell Avenue, a four-lane undivided highway, northwest from the highways' partial cloverleaf interchange with their mainline U.S. Highways and US 29. The business routes meet the northern end of SR 128 (Mayflower Drive) and cross over Norfolk Southern Railway's Blue Ridge District. Just north of the railroad, US 501 Business and US 460 Business veer onto Kemper Avenue, which crosses another rail line and has a cloverleaf interchange with US 29 Business (Lynchburg Expressway); this interchange serves as the northern terminus of US 221. The business routes and US 221 pass through a commercial area and by the Anne Spencer House, then turn west onto 12th Street and cross Norfolk Southern's Danville District rail line a few blocks south of the Lynchburg-Kemper Street Station, which is served by Amtrak.

At Campbell Avenue, US 221 and US 460 Business continue southwest on 12th Street while US 501 Business turns northwest onto Campbell Avenue, which becomes Langhorne Road after the intersection with Park Avenue. The business route intersects SR 163 (Memorial Avenue) at the historic home Centerview. West of Holy Cross Regional Catholic School, US 501 Business leaves the densely-populated part of Lynchburg and curves north, passing under a former railroad bridge. West of the Rivermont Historic District, the business route veers onto Rivermont Terrace, then turns west onto Rivermont Avenue, which becomes Boonsboro Road at its intersection with Link Road. US 501 Business continues on Boonsboro Road to its northern terminus at US 501, which heads south on the Lynchburg Expressway and north on the continuation of Boonsboro Road.

U.S. Route 501 Business (US 501 Business) is a business route of US 501 in Buena Vista. The highway runs 0.99 miles (1.59 km) between junctions with US 501 within the city of Buena Vista. When US 501 veers from Magnolia Avenue onto Sycamore Avenue at 16th Street, US 501 Business continues north on Magnolia Avenue, which serves as the main business street of the city. Just south of the Southern Virginia University campus, the business route veers northwest on Park Avenue, which the highway follows to its northern terminus at US 501 (Beech Street) just south of US 501's northern terminus at US 60.