The 58-year-old entertainer will play two dates in Los Angeles and two in New York to bring down the curtain on a live career that has spanned five decades.
Her manager, Martin Erlichman, said Streisand had "chosen to conclude her public performance career in the two cities most closely associated with her work".
She will give two farewell concerts at the Staples Centre in Los Angeles on 20 and 21 September and two more at New York's Madison Square Garden on the 27th and 28th.
Streisand, who is also a successful actress, has fought a lifelong battle with anxiety over singing live in public.
Many believed she would bow out after her sell-out millennium eve concert at the MGM Grand Hotel in Las Vegas, for which she was reportedly paid $5m (£3.3m).
"She thought that performance would be her last one," admitted publicist Dick Guttman. "But there were so many pressures from fans and others to conclude her career in New York and LA."
In addition to the September gigs, Guttman said Streisand would also be singing at a fund-raising concert for America's Democratic Party at the Shrine Auditorium in Los Angeles next month.
The Grammy-winning performer began her live singing career in the 1960s, but appearances since then have been few and far between.
Fear
Guttman said that prior to a pair of New Year concerts in 1993/94, she had not been paid to perform in front of an audience for 27 years.
"She acknowledged when she returned in '93 that she was overcoming a fear of public performance," he added.
Despite her aversion to live shows, her popularity has remained undimmed among her devoted fans.
With 42 gold-certified albums - 25 of which went platinum and 13 multi-platinum - Streisand is ranked as the biggest-selling female artist of all time by the Recording Industry of America.
She has had a string of hit singles in the UK - including the chart-topping Woman In Love and 1970s favourites You Don't Bring Me Flowers and No More Tears (Enough Is Enough).
In 1997 she enjoyed a futher two top ten hits, teaming up with Bryan Adams on I Finally Found Someone and Celine Dion for Tell Him.
Streisand's acting career has also proved to be a huge success. She made an Oscar-winning movie debut as Fanny Brice in 1968's Funny Girl and was widely praised for 1991's The Prince of Tides, in which she starred and directed.
In recent years she has been concentrating on her work as a TV and film producer.
Tickets for her farewell concerts go on sale on 30 July in Los Angeles and 31 July in New York.