Ferrari's Fernando Alonso won the 2010 Italian Grand Prix at Monza
Rome's Mayor Gianni Alemanno says the city has given up its ambition of hosting a Formula 1 race.
The announcement comes a week after F1 boss Bernie Ecclestone told Alemanno it would be difficult for Italy to stage more than one Grand Prix per year.
The Italian Grand Prix at Monza is already a fixture on the schedule.
At a news conference on Friday, Alemanno said: "Today we're definitively and formally renouncing the plan for Formula 1 in Rome."
Rome's project boss Maurizio Flammini had wanted a race through Rome's streets from 2013 and claimed that Ecclestone had proposed alternating the Italian GP between Rome and Monza.
However, Monza officials have said they are not prepared to share the hosting rights.
The Italian race will remain at Monza until 2016 after a deal between Ecclestone and the historic track's managers in March.
The Autodromo Nazionale Monza is one of F1's most famous venues, having first hosted the Italian Grand Prix in 1922. Monza's popularity with fans and teams also makes it highly unlikely an alternate hosting deal would be agreed.
Stefano Domenicali, boss of Italian team Ferrari, said that with 20 races now on the global calendar including newcomers India, he understood why there was no room for two in the same country.
And Alemanno, whose city is also bidding for the 2020 Olympics, was similarly realistic, saying last week: "If there must be a choice between the two races we will take a step back."
The current exception is Spain, which holds the Spanish Grand Prix in Barcelona and the European Grand Prix in Valencia, but the latter is set to rotate to other nations in the future as F1 expands to new markets.
The first Indian GP is due to take place in Greater Noida on the outskirts of Delhi on 30 October 2011, while South Korea hosted its first Grand Prix in 2010.