His father, the actor Robert Earl Jones, left his family before James Earl Jones was born, and the youth was raised largely by his grandparents in Michigan. He attended the University of Michigan (B.A., 1953), majoring in drama, and, after a brief stint in the U.S. Army, went to New York City, studying at the American Theatre Wing with Lee Strasberg (Strasberg, Lee). He acted in his first Off-Broadway production in 1957 and subsequently with the New York Shakespeare Festival in 1961–73. He won a Tony Award for his boxer role in Howard Sackler's The Great White Hope (1968) and later starred in the film version (1970). He received critical acclaim for the two-character stage play Paul Robeson (Robeson, Paul) (1978) and in the title role of Othello, opposite Christopher Plummer's Iago (1981–82).

A part in the film Dr. Strangelove (1964) began a prolific career in pictures for Jones, whose roles included an evil ruler in the fantasy film Conan the Barbarian (1982), a coal miner fighting for the right to form a union in Matewan (1987), and an African king who lets his son (played by Eddie Murphy (Murphy, Eddie)) travel to the United States in the comedy Coming to America (1988). He appeared as Admiral James Greer in the film adaptations of Tom Clancy's novels about CIA agent Jack Ryan: The Hunt for Red October (1990), Patriot Games (1992), and Clear and Present Danger (1994). In 1995 he portrayed Rev. Stephen Kumalo in the film version of Alan Paton (Paton, Alan)'s classic novel Cry, the Beloved Country. Jones next starred opposite Robert Duvall (Duvall, Robert) in A Family Thing (1996). In 2008 he appeared as Big Daddy in a Broadway production of Tennessee Williams's Cat on a Hot Tin Roof that featured an all-black cast.

Known for his deep, resonant voice, Jones was cast in many voice-over roles in television advertising and in films, both as a narrator and for animated characters. He is perhaps best known for giving voice to the villain Darth Vader in the Star Wars series of movies, which began in 1977. In 1994 he provided the voice of Mufasa in Disney's The Lion King.