Attached to the 1st U-boat Flotilla, she made nine successful patrols in the North Atlantic, the last two under the command of Oberleutnant zur See Günther Rosenberg.

The submarine was powered by two Germaniawerft F46 four-stroke, six-cylinder supercharged diesel engines producing a total of 2,800 to 3,200 metric horsepower (2,060 to 2,350 kW; 2,760 to 3,160 shp) for use while surfaced, two AEG GU 460/8–27 double-acting electric motors producing a total of 750 metric horsepower (550 kW; 740 shp) for use while submerged.

Her route took her across the North Sea, through the gap separating Iceland and the Faroe Islands and into the Atlantic Ocean.

The submarine's second foray passed without major incident: starting on 8 June 1941, finishing on 19 July but in Brest.

Firing one spread of four torpedoes she hit the cargo ship Ciscar and passenger liner Aguila, which was carrying the Convoy Commodore and 86 other Royal Navy personnel.

U-201 continued with the concerted attack on OG 71, sinking the Irish Clonlara on 22 August and British merchants Aldergrove and Stork northwest of Lisbon on the 23rd, before returning to Brest on the 25th.

She also accounted for HMS Springbank, a Fighter catapult ship about 430 nmi (800 km; 490 mi) west southwest of Cape Clear, southern Ireland on the same date.

Having departed Brest and crossed the Atlantic, she damaged the Argentinian and neutral Victoria about 300 nmi (560 km; 350 mi) east of Cape Hatteras, North Carolina on 18 April.

Victoria's complement then abandoned its vessel; U-201 reported its mistake to the BdU (U-boat headquarters) which ordered it to clear the area, which it did.

USS Owl, an American minesweeper towing the barge YOG-38, picked up Victoria's distress signals and sent a boarding party across to the tanker to effect repairs.

The ship reached New York on 21 April and after much legal wrangling, was repaired and requisitioned by the US government and returned to service in July.

The 48 survivors were spotted by a Yugoslavian merchant ship, but when they learned of the prospect of an unescorted Atlantic crossing to Durban, opted to remain in their lifeboats until they reached the mouth of the River Marowijine.

U-201 was sunk by depth charges from the British destroyer HMS Fame east of Newfoundland on 17 February 1943.