Financially-troubled US F1 will have to submit a new application to enter the 2011 Formula One world championship.
That is the decision of the FIA as motor sport's world governing body definitively confirmed this year's entry list that only includes 12 teams, not the 13 as planned.
It also means there is no place on the grid for Stefan GP who had been hoping to gain an entry, even at such a late hour, if any of the other teams fell by the wayside.
Confirming the position, a FIA statement read: "The US F1 Team have indicated they will not be in a position to participate in the 2010 FIA Formula One world championship."
"Having considered the various options, the FIA confirms it is not possible for a replacement team to be entered for the championship at this late stage."
"In the coming days the FIA will announce details of a new selection process to identify candidates to fill any vacancies existing at the start of the 2011 season."
Given its financial woes, US F1 had asked the FIA to defer its entry until next season, but the sport's authority have made clear that is not guaranteed and it will again be subject to a bidding process.
US F1 team principal Ken Anderson had hoped the FIA would take pity on his marque's plight and give them the year's grace.
Given the FIA's decision, it now remains to be seen whether US F1 will soldier on as Anderson had ordered his staff to halt work on the car until an announcement was made.
US F1's problems started a few months ago after sponsors started to pull out, which led to production on the car grinding to a halt.
The team have since struggled to pay wages on time, whilst several members of staff have chosen to leave.
Despite that, US F1 still announced the signing of its first driver at the end of January in Argentinian Jose Maria Lopez.
Lopez, financed by 5million pounds worth of sponsorship, is now without a drive for this year, although he appears likely to be handed the test and reserve role with Hispania Racing, formerly Campos Meta.
Stefan GP, run by Serbian businessman Zoran Stefanovic and who are based in Toyota's old headquarters in Cologne, had been waiting in the wings in the hope of being granted an 11th-hour slot.
Stefanovic had an agreement to purchase the chassis as Toyota had already put into production their 2010 model prior to the shock announcement at the end of last season they were quitting F1.
Stefan GP was leasing an engine, gearbox, race team equipment and trucks, as well as office and workshop space in the former Toyota factory.
A number of drivers, including former champion Jacques Villeneuve, had held talks with Stefanovic in the hope they would be racing this year.
But as with US F1, Stefan GP will now have to contest the place on the grid for 2011.
The FIA's decision opens the door for the likes of David Richards' Prodrive organisation to again re-apply as they were one of the unsuccessful applicants last year.