Just four months after being appointed, the decision has been taken to end the Portuguese's tenure following a poor run of results.
He got off to a flyer with a maximum nine points from the opening three Premier League matches of the season, putting Spurs top of the table going into the September international break.
However, Spurs' form has plummeted since with five defeats in their last seven league outings.
Spurs' 3-0 humbling at home to Manchester United saw fans at the Tottenham Hotspur Stadium boo Nuno's decision to substitute Lucas Moura and chant 'you don't know what you're doing'.
There were also chants of 'Levy out' during the match as supporters turned on the chairman, as well as striker Harry Kane, too.
A Tottenham statement published on Monday morning read: "The club can today announce that Nuno Espirito Santo and his coaching staff Ian Cathro, Rui Barbosa and Antonio Dias have been relieved of their duties.
"A further coaching update will follow in due course."
Fabio Paratici, managing director of football, said: "I know how much Nuno and his coaching staff wanted to succeed and I regret that we have had to take this decision.
"Nuno is a true gentleman and will always be welcome here. We should like to thank him and his coaching staff and wish them well for the future."
Chairman Daniel Levy and Paratici now have the task of finding a permanent successor to Nuno.
A number of big names have been linked to the post, with former Chelsea boss Antonio Conte the bookies favourite, while Brighton boss Graham Potter is second favourite.
Reported Newcastle target Paulo Fonseca, who was understood to have been interviewed for the Spurs job before Nuno got it, is also in the running as are club legend Jurgen Klinsmann and former manager Mauricio Pochettino.
Below you can see the contenders to be Tottenham's next head coach and what odds you can get from Betfair.