Stars from Hollywood, Nashville and Broadway gathered in the nation's capital to salute Winfrey, McCartney and three others - country singer Merle Haggard, Broadway composer Jerry Herman and dancer Bill T. Jones - with the Kennedy Center Honors.
President Obama and first lady Michelle Obama sat with the honorees and former Secretary of State Colin Powell.
Julia Roberts opened the show with a surprise nod to her friend, Winfrey.
"It's a universal conversation starter: Did you see what was on 'Oprah' today?" Roberts said. "The first time I heard of a better fitting bra ... or a fascinating politician named Barack Obama was on 'The Oprah Winfrey Show."'
Roberts said she was nervous to address the Washington crowd but nearly tricked them into looking under their seats for a prize - Winfrey style.
John Travolta took the stage to host a mock version of Winfrey's show with Barbara Walters as his guest. He recounted a phone call he got from Winfrey when Hurricane Katrina struck New Orleans. She told him to fill his plane with medical supplies and meet her in Louisiana.
"So when Oprah calls, you answer," he said. "Oprah makes it exciting to be responsible."
Walters said she should get credit for jump-starting the 56-year-old Winfrey's career. She said the future talk show host once imitated Walters' style to win a Miss Fire Prevention contest but developed a style all her own.
"Simply put, she is the best interviewer ever," Walters said. "No one comes close - not even me. And those of you who know me know how painful it was for me to say that."
Comedian Chris Rock roasted Winfrey, saying, "No one deserves this award more than Oprah Winfrey, but no one needs it less!"
Jennifer Hudson sang "I'm Here" from "The Color Purple," which Winfrey produced on Broadway after starring in the film. Hudson was joined by a choir from Winfrey's alma mater, Tennessee State University.
For Winfrey, the prize comes during the 25th and final season of her talk show and just before she launches her new cable network, OWN, on Jan. 1. After her Washington visit, Winfrey will take about 300 audience members to Australia.
Winfrey told "Early Show" co-anchor Julie Chen, who also co-hosts the CBS show "The Talk," that she understands the immensity of the honor, saying, "I've sat in auditoriums and saw all of the names up on the stage, going, going, 'Whoa, how impressive!' … so I understood deeply what it meant to get this award."
Winfrey's best friend Gayle King told The Associated Press it was a fitting tribute for Winfrey, saying, "What's interesting is she spends her life celebrating others, but when it comes time for her, she's very reluctant really. .. They're recognizing her whole body of work. She's not just a talk show host."
Alec Baldwin introduced the tribute for McCartney, 68, lamenting the singer's "long and winding road" to a solo career, being forced to sing in stadiums and requiring police protection.
"The National Institutes of Health called the epidemic Beatlemania," Baldwin said. "There was no cure."
The former Beatle was making his second visit to Washington this year for a culture award. In June, he won the Gershwin Prize for Popular Song from the Library of Congress.
"You know, great things just come in bundles," he said.
Gwen Stefani, dressed in a gray "Fab Four-style" pant suit, and her band, No Doubt, opened the musical set with "Hello, Goodbye." Dave Grohl and Norah Jones sang "Maybe I'm Amazed."
In what's perhaps a first, Aerosmith front man Steven Tyler performed an "Abbey Road" medley live. Show producer George Stevens Jr. said The Beatles never performed it live after the recording was made.
"I've done a lot of things and been a lot of places," Tyler said. "But I've never been prouder than I am standing right here at the corner of Pennsylvania Avenue and Abbey Road."
James Taylor and Mavis Staples closed out the show with "Let It Be" and "Hey Jude." They had McCartney - and the president - on their feet the entire time.
When The Beatles were storming America, Haggard, 73, was free from prison and making music, Herman, 79, was making Broadway sing and Jones, 58, was not yet a dancer but growing up in a migrant labor camp.
Since the 1960s, the new Kennedy Center honorees have helped define television, dance, theater and music.
At a White House ceremony before the show Sunday, they heard accolades from President Barack Obama who said the awards are important because the arts are necessary.
"Although the honorees on this stage each possess a staggering amount of talent, the truth is, they aren't being recognized tonight simply because of their careers as great lyricists or songwriters or dancers or entertainers," Obama said. "Instead, they're being honored for their unique ability to bring us closer together and to capture something larger about who we are - not just as Americans, but as human beings."
The show was recorded Sunday night for broadcast Dec. 28 on CBS.
At the gala performance, Angela Lansbury, Carol Channing and Kelsey Grammer performed some of Herman's famous tunes from "Hello, Dolly!," "Mame," and "La Cage aux Folles."
Lansbury said Herman gave her a huge break when he pressed for her to be cast in the lead role of "Mame" over the objections of some producers. She said she's never had a better entrance than sliding down a long banister on stage with bugle in hand.
Channing, 89, who starred in "Hello, Dolly!" sang "Hello, Jerry." She said she knew it would make him cry.
For Haggard, Nashville turned out in force.
Willie Nelson and Sheryl Crow sang Haggard's "Today I Started Loving You Again," and Jamey Johnson and Kid Rock sang "Ramblin Fever."
Vince Gill and Brad Paisley sang "Workin Man Blues."
After the honors were announced in September, Jones, the son of potato pickers, said he could recall dreaming of big things as a 9-year-old boy in upstate New York.
He went on to create the Bill T. Jones/Arnie Zane Dance Company in 1982 with his partner Arnie Zane who died of AIDS six years later. Jones has tackled tough issues with his work, such as racism and mortality, sometimes sparking outrage.
On Sunday, he was hailed as a rule breaker and revolutionary by playwright Edward Albee, Claire Danes and others.
Jones said he's often felt like an outsider, yet he's being honored for helping to shape the country.
Jones said he could "feel the love" and said he was thrilled to be receiving the award while Obama is president. Still, he said he's discouraged by the country's direction. "I am trembling a little bit, actually. The discourse is so poisonous."
In recent years, Jones has shared his talents with Broadway, winning Tony awards for his choreography in "Spring Awakening" and this year for his show "Fela!," playing currently in New York and London. He said he hopes more artists will bridge the gap between the dance world and mainstream theater.
"We don't want to be marginalized anymore," he said. "Indifference is worse than dislike."