Holmes, who earlier this month reached a high-profile divorce agreement with "Mission: Impossible" actor Cruise, will star in Theresa Rebeck's new comedy, representatives for the show said in a statement.
The actress will portray the role of Lorna in the five-character play that tackles "issues of corporate greed, small-town values and whether or not your family will always welcome you back ... with no questions asked," the statement said.
"I am thrilled to be coming back to the Broadway community and honoured to be a part of this team," Holmes said in a statement to the New York Times.
The play will be directed by Tony Award winner Jack O'Brien.
Holmes, 33, made her Broadway debut in the 2008 production of Arthur Miller's "All My Sons," and she recently moved to New York City with her and Cruise's 6-year-old daughter, Suri.
The former "Dawson's Creek" actress and Cruise, 50, took less than two weeks to end a nearly six-year marriage that had been a favourite subject for celebrity magazines.
During the marriage, Holmes appeared in several TV movies and shows, including the miniseries "The Kennedys." However her career seemed overshadowed by Cruise, who saw a resurgence with recent box office hit, "Mission: Impossible - Ghost Protocol."
Last week, Cruise visited Suri in New York for the first time since the divorce settlement after he had been filming in California and Iceland.
