Assigned to the Atlantic Fleet, Reuben James sailed from Newport, Rhode Island, 30 November 1920 to Zelenika, Yugoslavia, arriving 18 December. During the spring and summer of 1921, she operated in the Adriatic and the Mediterranean out of Zelenika and Gruz, Yugoslavia, assisting refugees and participating in postwar investigations. In October 1921 at Le Havre, she joined Olympia (C 6) at ceremonies marking the return of the Unknown Soldier to the United States. At Danzig, Poland, from 29 October 1921 to 3 February 1922, she assisted the American Relief Administration in its efforts to relieve hunger and misery. After duty in the Mediterranean, she departed Gibraltar 17 July 1922.

Based then at New York, she patrolled the Nicaraguan coast to prevent the delivery of weapons to revolutionaries in early 1926. In the spring of 1929, she participated in fleet maneuvers that foreshadowed naval airpower. Reuben James decommissioned at Philadelphia on 20 January 1931.

Recommissioned 9 March 1932, she again operated in the Atlantic and the Caribbean. From September 1933 to January 1934 she patrolled Cuban waters during a period of revolution. Sailing for the Pacific from Norfolk 19 October 1934, she arrived at her new homeport of San Diego, Calif., 9 November. Following maneuvers that evaluated aircraft carriers, she returned to the Atlantic Fleet in January 1939. Upon the outbreak of war in Europe in September 1939, she joined the Neutrality Patrol, and guarded the Atlantic and Caribbean approaches to the American coast.

In March 1941, Reuben James joined the convoy escort force established to promote the safe arrival of war materials to Britain. This escort force guarded convoys as far as Iceland, where they became the responsibility of British escorts. Based at Hvalfjordur, Iceland, she sailed from Argentia, Newfoundland, 23 October 1941, with four other destroyers to escort eastbound convoy HX-156. While escorting that convoy, at about 0525 on 31 October 1941, Reuben James was torpedoed by German submarine U-562. Her magazine exploded, and she sank quickly. Of the crew, 44 survived and 115 died.

Reuben James was the first US Navy ship sunk by hostile action in World War II.