Winfrey's first name was spelled "Orpah" on her birth certificate after the biblical figure in the Book of Ruth, but people mispronounced it regularly and "Oprah" stuck. She was born in Kosciusko, Mississippi, to an unmarried teenage mother. She later said that her conception was due to a single sexual encounter and the couple broke up not long after. Her mother, Vernita Lee (born c. 1935), was a housemaid.

In 1985, Winfrey co-starred in Steven Spielberg's The Color Purple as distraught housewife Sofia. She was nominated for an Academy Award for Best Supporting Actress for her performance. The Alice Walker novel went on to become a Broadway musical which opened in late 2005, with Winfrey credited as a producer. In October 1998, Winfrey produced and starred in the film Beloved, based on Toni Morrison's Pulitzer Prize-winning novel of the same name. To prepare for her role as Sethe, the protagonist and former slave, Winfrey experienced a 24-hour simulation of the experience of slavery, which included being tied up and blindfolded and left alone in the woods. Despite major advertising, including two episodes of her talk show dedicated solely to the film, and moderate to good critical reviews, Beloved opened to poor box-office results, losing approximately $30 million. While promoting the movie, co-star Thandie Newton described Winfrey as "a very strong technical actress and it's because she's so smart. She's acute. She's got a mind like a razor blade." In 2005, Harpo Productions released a film adaptation of Zora Neale Hurston's 1937 novel Their Eyes Were Watching God. The made-for-television film was based upon a teleplay by Suzan-Lori Parks and starred Halle Berry in the lead female role.  In late 2008, Winfrey's company Harpo Films signed an exclusive output pact to develop and produce scripted series, documentaries, and movies for HBO.  Oprah voiced Gussie the goose in Charlotte's Web (2006) and voiced Judge Bumbleton in Bee Movie (2007), co-starring the voices of Jerry Seinfeld and Renee Zellweger. In 2009, Winfrey provided the voice for the character of Eudora, the mother of Princess Tiana, in Disney's The Princess and the Frog and in 2010, narrated the US version of the BBC nature program Life for Discovery.  In 2018, Winfrey starred as Mrs. Which in the film adaptation of Madeleine L'Engle's novel A Wrinkle in Time.

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