Worcester Warriors have been suspended from all competitions after failing to meet a funding ultimatum set by the Rugby Football Union (RFU).
The financially stricken club had to provide proof of a "credible" plan for the future by 17:00 BST on Monday.
The men's team will now be banned from the Premiership and its women's side from the Premier 15s.
"We have had to take this action to protect everyone's best interests," RFU chief executive Bill Sweeney said.
Sweeney added that the news was "incredibly difficult for fans, staff and players" and thanked them for working "tirelessly over recent weeks to enable matches to continue".
What happens next to Warriors is not confirmed - with the club yet to enter administration - but Sweeney hopes a buyer can be found as soon as possible, with the club potentially able to resume matches if the suspension is lifted.
The RFU are yet to announce any further sanctions on Warriors, which may include a 35-point deduction or possible relegation.
"While it is the responsibility of each business owner to manage their individual finances, we will look at learnings from this situation to see what regulation can be put in place to provide all parties with more financial transparency.
"Rugby is a relatively young professional sport and it has been widely recognised that clubs have been facing financial challenges even before Covid.
"Successful professional leagues are vital for the wellbeing of the whole game. This is why it is so important that we continue to work with Premiership Rugby to improve the structure, governance and business model of rugby union in England."
The ongoing Worcester saga
Following Worcester's suspension, Premiership Rugby have confirmed their scheduled Premiership match at Gloucester will not now go ahead, adding it will "continue to support them where we can during this next phase".
The Cherry and Whites tweeted: "Our thoughts are with the Worcester Warriors players, staff and supporters. We hope to see them return to the Premiership again soon."
Warriors are not the only club struggling financially, with fellow Midlands side Wasps announcing their intention to appoint administrators to "protect the club's interests".
"Wasps and the entire rugby family stands as one with all the players, staff and supporters of Worcester Warriors," the club tweeted after Worcester's suspension was confirmed.
The club's failure to meet the RFU's deadline was not unexpected, with staff told in a letter to collect their belongings by 16:30 BST, when the gates to Sixways Stadium would be locked, with the club's insurance policy expiring at midnight.
With access to the stadium and its facilities no longer available, the players are now set to train on local pitches while their futures are sorted out, with caretaker Lee Morrow and his wife having to leave their flat above the West Stand.
Despite the two previous threats of suspension, Warriors staged their first two home games of the Premiership season as many staff worked for free, with capacity at Sixways reduced to 4,999 due to safety regulations.
Those that went on Saturday saw Warriors win an emotionally charged match 39-5, while the club's women's side - the University of Worcester Warriors - have won both their first two matches in the Allianz Cup.
Warriors ban follows weeks of financial turmoil ever since they were served with a winding-up petition by His Majesty's Revenue & Customs over a £6m unpaid tax bill.
The club were also given a loan through Sport England, thought to be around £15m, as part of the government's sports survival package (SSP) during the coronavirus pandemic. The club's total debts are believed to be £25m.
Players were paid late and staff only partially paid their salaries for August with others not receiving any at all.
Despite assurances from owners Jason Whittingham and Colin Goldring, the rest of those salaries - 35% - went unpaid and no official confirmation ever followed their announcement of an agreement to sell the club nearly two weeks ago.
The owners have so far rejected calls from local MPs to put the club into administration, prompting Worcester MP Robin Walker to ask the Department for Culture, Media and Sport (DCMS) in parliament last week to trigger the administration process.
The closure of Sixways has also had a knock-on effect for non-league football club Worcester Raiders, who play their home matches at the stadium.