David Dukes was the eldest of four boys: David, James, Robert and Joe Paul. He married his first wife while a student at the College of Marin on October 9, 1965. Their son Shawn David Dukes was born on March 31, 1966.

Dukes had considerable stage experience, first appearing on Broadway in 1971. He later appeared in a revival of Molière's The School for Wives. Dukes' theatrical roles included as Dracula, Doctor Frankenstein, and Antonio Salieri in the original production of Amadeus, replacing Ian McKellen. He also replaced John Lithgow in the original production of David Henry Hwang's play M. Butterfly, and he received a Tony nomination in 1980 for best featured actor in a play for Bent.

Dukes' film career included 35 movies. Throughout his career, he was a television guest star, notably as the man who attempted to rape Edith Bunker on All in the Family and as a blind bully on Three's Company. During the 1980s, Dukes appeared in the dual miniseries The Winds of War and War and Remembrance. He received an Emmy nomination for best supporting actor for his role in The Josephine Baker Story (1991) and appeared as Arthur Miller in Norma Jean & Marilyn (1996). He was a regular in the first season of Sisters, playing the transvestite husband of oldest sister, Alex (Swoosie Kurtz). Dukes' role became a recurring character in subsequent seasons. On Dawson's Creek, he had the recurring role of Mr. McPhee, father to Jack (Kerr Smith) and Andie (Meredith Monroe) from the second through fourth seasons. He also starred in Without A Trace as the ex-husband of Kate Nelligan.