Juventus have been told they face expulsion from Serie A if they remain part of the European Super League.
The Turin club, along with Barcelona and Real Madrid, are yet to withdraw from the proposed breakaway league.
Italian federation president Gabriele Gravina said: "If Juventus does not respect the rules they will be out.
"At the time of registration for the next Serie A championship the club will be excluded if they decide not to withdraw from the Super League."
Juve, Barca and Real continue to be committed to the project, and last week defended their involvement in the breakaway league in the face of what they call "threats" from Uefa.
But pressure is mounting on Serie A club Juve to withdraw.
"We are all a bit tired of this tug-of-war between Uefa and these three clubs," said Gravina. "I hope this dispute can be resolved as soon as possible. I hope to be able to mediate between Juventus and Uefa.
"The rules foresee the non-participation in our championship if the principles established by the federation and Uefa are not accepted.
"It's not good for international football, Italian football, Juventus. We have already said that the football association respects the rules."
The ESL was announced on 18 April but within 48 hours the plans had fallen apart with the English clubs withdrawing after fan protests and UK government pressure.
The nine clubs who withdrew from the competition have been given a financial punishment by Uefa.
Manchester United, Manchester City, Liverpool, Chelsea, Tottenham and Arsenal, along with AC Milan, Inter Milan and Atletico Madrid, also committed to the European governing body and its competitions.
They have agreed to make a combined 15m euro (£13.4m) goodwill contribution to benefit children's and grassroots football across Europe.
- Snowfall is back: Hard-hitting drama about the crack epidemic in 1980s LA streaming now
- How Gen Z is learning to adapt and thrive: Young entrepreneurs find new ways to make money in Covid times