The 47-year-old Portuguese was appointed Spurs manager in the summer but his time in charge of the North Londoners lasted just 17 games.
Esperito Santo had a bright start to his Tottenham tenure, winning his opening three games to be crowned Premier League manager of the month for August.
Spurs have however lost five of their subsequent seven top flight games and they lie eighth on the table with 15 points.
A 3-0 defeat to Manchester United on Saturday has proven to be the former Wolverhampton Wanderers manager's final match in charge.
An official statement on the Spurs website read: "The Club can today announce that Nuno Espírito Santo and his coaching staff Ian Cathro, Rui Barbosa and Antonio Dias have been relieved of their duties."
Fabio Paratici, Managing Director of Football, added: "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.”
Esperito Santo is the third Premier League manager to be sacked this season after Watford parted company with Xisco Munoz and Newcastle United dismissed Steve Bruce.
Former Juventus, Chelsea and Inter Milan manager Antonio Conte is the early front-runner to be named as the new Spurs manager.
Tottenham's next league outing is against Everton this weekend.
