Following shakedown training which took Jouett to England and Ireland, the ship returned to Norfolk 29 April 1939 and began operating on neutrality patrol along the East and Gulf Coasts. She stood out of Pensacola Bay 15 February 1940 as one of the escorts for Tuscaloosa (CA 37), carrying President Roosevelt on a cruise through the Gulf of Panama, returning to Pensacola 1 March 1940. Jouett then set course for the Panama Canal and the Pacific, arriving Pearl Harbor for duty 10 April 1940.
The destroyer remained in Hawaiian waters during the next year exercising with America's vital carriers and perfecting tactics. Sailing 18 April 1941, Jouett accompanied Yorktown (CV 5) through the canal to Cuba, proceeding from there to Port-of-Spain, Trinidad, 19 May. The ship then joined a cruiser and destroyer force under Rear Admiral Jonas H. Ingram charged with guarding against German surface or submarine attacks on American shipping. Jouett was at Port of Spain 7 December 1941 when the Japanese attack on Pearl Harbor brought America into the war. The ship then began offensive antisubmarine patrols between Brazil and Africa, helping to keep the all-important ocean supply lines open. She accompanied Army engineers to lonely Ascension Island 30 March 1942 where an airfield was curved from the bleak landscape. Jouett convoyed the precious oil tankers from Trinidad south during the months that followed, often attacking submarines with depth charges.
In December 1942, the ship returned to Charleston for repairs, but by 21 January 1943 she was back in Natal harbor, Brazil. Jouett received President Vargas of Brazil 27 January 1943, providing quarters for him and his party during important conferences on board Humbolt (AG 121) with President Roosevelt. Following the talks, which cemented relations between the countries and provided for closer naval cooperation, President Vargas departed Jouett 29 January.
The veteran destroyer resumed her escort duties in February, and 14 May joined in the search for U-128 off Bahia, Brazil. Aircraft dropped depth charges on the sub and brought her to the surface where gunfire from Jouett and Moffett (DD 362) sent her to the bottom. The destroyer continued to serve with Admiral Ingram’s crack antisubmarine force, now Fourth Fleet, through the rest of 1943. On New Year’s Day 1944 she joined Omaha (CL 4) for ocean patrol; and the ships intercepted German blockade runner SS Rio Grande, with a vital cargo of crude rubber. After the crew abandoned ship, Omaha and Jouett sank the German ship. This effective closing of the South Atlantic to German blockade runners was demonstrated even more forcefully 5 January when patrol planes reported a strange ship identifying herself as Flordian. Intelligence identified her, however, as blockade runner Burgenland. Before aerial attacks could begin Omaha and Jouett picked her up on radar and closed in. Scuttling charges and the cruiser's gunfire sank her just after 1730.
Jouett returned to Charleston once more in March 1944 and engaged in training operations in Casco Bay, Maine, before sailing for England in convoy 16 May 1944. There she joined a Reserve Fire Support Group for the long awaited invasion of France. Jouett arrived off Omaha beach 8 June, escorting coastal steamers with support troops embarked. She repelled an air attack that day, and until 21 June screened British heavies during shore bombardment and provided antisubmarine screen for the Omaha Beach transport area. The second front established, Jouett escorted convoys to and from the Firth of Clyde until 12 July 1944 when she sailed with a convoy for Algeria.
The destroyer arrived at Oran 21 July to prepare for the next major European operation, the invasion of southern France. Departing Naples 14 August, Jouett arrived off the Delta assault area next day and. as troops landed, acted as command ship of the Convoy Control Group charged with the smooth routing and unloading of support troops. This duty continued until 3 September, after which the ship operated on patrol out of Toulon. In early October Jouett steamed off Cape Ferrat, giving gunfire support to American troops in the bitter fighting ashore. She also destroyed mines off San Remo 9 October, destroyed bridges, and covered Allied minesweeping operations in the area.
Jouett sailed from Oran 31 December 1944 for repairs at Charleston. After refresher training in Casco Bay in April, the battle tested ship made convoy voyages to England and Cuba before the end of the war 15 August 1945. She decommissioned at Philadelphia Naval Shipyard 1 November 1945 and was scrapped there in 1946.
Jouett received three battle stars for World War II service.