Sutherland was born in Sydney. In 1951, she won Australia's most important vocal competition and went to London to study at the Opera School of the Royal College of Music. She made her European debut at the Royal Opera House at Covent Garden on 28 October 1952 as the First Lady in Die Zauberflöte.
Sutherland decided to concentrate on the bel canto repertoire and her major breakthrough was the famous 1959 production of Lucia di Lammermoor conducted by Tullio Serafin and staged by Franco Zeffirelli.
She went on to sing Lucia to great acclaim in Paris the following year, and at La Scala and the Metropolitan Opera in 1961. An accomplished exponent of Handel, she sang the title role in Alcina for her debut at La Fenice, Venice in 1960 and at Dallas, with which she made her US debut the same year.
She later expanded her repertoire to include Amina (La Sonnambula) , Violetta (La Traviata) and Elvira (I Puritani) alongside Semiramide, Marguerite de Valois (Les Huguenots), Marguerite, Lakme, Cleopatra (Giulio Cesare) and Norma. She was particularly admired for her performance of Marie, the tomboy soldier in La Fille du Regiment.
Later in her career, she added roles such as Tales of Hoffmann (singing all four roles), Lucrezia Borgia, Anna Bolena , Esclarmonde, and Adriana Lecouvreur.
Sutherland is married to conductor Richard Bonynge, with whom she performed many times and made many recordings.