She debuted on Broadway in 1957 and, in 1975, won a Tony Award for Best Actress in a Play for her performance in "Same Time, Next Year." In 1990 she won the Sarah Siddons Award for her work in Chicago theatre.

Burstyn received her first Oscar nomination for Best Supporting Actress in 1972 for the film The Last Picture Show. She subsequently won the Best Actress Oscar in 1974 for her performance in the movie Alice Doesn't Live Here Anymore and was nominated for Best Actress in 1973 for the horror movie The Exorcist, in 1978 for Same Time, Next Year, and in 1980 for Resurrection. She also received an Oscar nomination for her role as Sara Goldfarb in Requiem for a Dream in 2000, a performance that is generally considered to be the finest of her career and one of the best of the decade. However, Julia Roberts won the Oscar for Best Actress for Erin Brokovich in 2000. In retrospect, many film critics decried the choice as pandering to popular demand over an objective evaluation of performance. From 2000 to 2002, Burnstyn appeared the CBS television drama That's Life.

Burstyn served as president of the Actors' Equity Association from 1982 and 1985.

In 1997, Burstyn was inducted into the Michigan Women's Hall of Fame.

In 2000 Burstyn was, along with Al Pacino and Harvey Keitel, named co-president of The Actor's Studio. In 2004, Burstyn was confirmed to play a key role as Lilian in Darren Aronofsky's upcoming The Fountain.