Tuttle was born August 29, 1907, at Pleasant Lake, Indiana, into a family with strong ties to entertainment. Her father, O.V. Tuttle, had been a performer in minstrel shows before becoming a station agent for a railroad. Her grandfather, Frank Tuttle, managed an opera house and taught drama. She discovered her own knack for acting after moving with her family to Glendale, Arizona. She later credited a drama coach there for "making me aware of life as it really is—by making me study life in real situations."

Tuttle had a hobby of collecting toy dogs. A 1930 newspaper article reported, "Her dressing room shelf is filled with more than 200 miniature replicas of every variety of dog known."

Tuttle's radio debut came in 1936 when she appeared on Hollywood Hotel with Dick Powell. Despite having never performed before a microphone, Tuttle's audition won her a three-year contract with the program.

Tuttle played the swallow in "The Happy Prince", an adaption of Oscar Wilde's short story with Orson Welles and Bing Crosby (1946). The story had earlier been adapted for radio by Orson Welles in 1944, featuring a musical score by Bernard Herrmann. It aired on the Philco Radio Hall of Fame broadcast on December 24, 1944 with Lureen Tuttle playing The Swallow and featuring Bing Crosby alongside Orson Welles, with Herrmann's music conducted by Victor Young.

Heaven Only Knows (1947) was her first film. She went on to roles in other films such as Orson Welles's Macbeth (1948), Mr. Blandings Builds His Dream House (1948) and Alfred Hitchcock's Psycho (1960), as the wife of Sheriff Chambers. In Don't Bother to Knock (1952) she portrayed a mother who lets a disturbed Marilyn Monroe babysit her daughter. The next year she appeared again with Marilyn in Niagara, as Mrs. Kettering. She had a rare starring role in Ma Barker's Killer Brood (1960). She played Grandma Pusser in the original Walking Tall film trilogy, and also appeared in horror films such as The Manitou (1978), starring Tony Curtis. Her final film role was in the 1983 film Testament.

Tuttle married Melville Ruick, an actor whom she had met during her radio years; the couple had a daughter, Barbara Ruick,. Tuttle and Ruick eventually divorced. She married Frederick W. Cole, an Engineer, on November 27, 1950 in Pasadena, California; she sued him for divorce on January 4, 1956.

In 1958 and 1959, she was cast in two episodes as Gladys Purvis, the mother of series character Kate McCoy, played by Kathleen Nolan, in the ABC sitcom, The Real McCoys, with Walter Brennan and Richard Crenna. She appeared twice on the NBC western series The Californians, once as Belle Calhoun in "Skeleton in the Closet" (1958) and then as Maude Sorel in "The Painted Lady" (1959). She guest-starred with Andrew Duggan in his crime series, Bourbon Street Beat.

Tuttle has two stars on the Hollywood Walk of Fame – "Star of Radio" at 1760 Vine Street and "Star of Television" at 7011 Hollywood Boulevard. Both stars were dedicated February 8, 1960.

Tuttle was cast as Mrs. Grange in the 1963 episode "The Risk" on the NBC drama series Mr. Novak, starring James Franciscus as an idealistic high school Teacher in Los Angeles. She later appeared on the popular 1960s sitcoms I Dream of Jeannie, The Munsters," and "Petticoat Junction".

Tuttle's best-known role to the general public was in 32 episodes of the NBC series Julia (1968–1971) as the humorless but warm-hearted Hannah Yarby.

In 1972, she appeared as Bella Swann, on the season three episode, "Farmer Ted and the News" on The Mary Tyler Moore Show.

In 1980, Tuttle appeared as Mrs. McIntyre in the television movie, White Mama, with Bette Davis. From 1981 to 1984, Tuttle appeared six times on the CBS medical drama series Trapper John, M.D..

Tuttle died from cancer May 28, 1986, at Encino Hospital. She was survived by a granddaughter, two grandsons, and a great-granddaughter. Memorial services were held June 2, 1986, at Church of the Recessional at Forest Lawn Memorial Park, Glendale.

In 1985, Tuttle appeared in episode 25 ("Murder in the Afternoon") of "Murder, She Wrote", playing the character Agnes Cochran, Jessica Fletcher's aunt. The episode first aired 10/13/85. [The Unofficial Murder She Wrote Casebook by James Robert Parish.]

She became a respected acting coach and teacher—something she had always done, even at the height of her acting career (she often re-trained radio actors who had been away from the craft during Service in World War II).