Andy Gray was sacked by Sky Sports today after footage emerged of the presenter appearing to make an inappropriate remark to a female colleague.
The footage came into the public domain two days after Gray was shown making controversial comments about assistant referee Sian Massey.
Gray had his contract terminated following the release of the new footage, 24 hours after he and fellow presenter Richard Keys were disciplined for their remarks about Massey.
Sky Sports reporter Andy Burton was also disciplined this afternoon after footage emerged of him and Gray commenting on Massey's appearance.
Announcing the departure of Gray, who has been one of the faces of the broadcaster's live Barclays Premier League coverage since it began almost 20 years ago, Sky Sports said in a statement: "Sky Sports today terminated its contract with Andy Gray.
"The decision, which is effective immediately, was made in response to new evidence of unacceptable and offensive behaviour.
"The new evidence, relating to an off-air incident that took place in December 2010, came to light after Andy Gray had already been subjected to disciplinary action for his comments of 22nd January 2011."
The new footage, which was recorded last month but only came to light in the last 24 hours, appears to show Gray making a suggestive comment towards colleague Charlotte Jackson, who does not openly react while he and Keys then burst out laughing.
Managing director Barney Francis added: "Andy Gray's contract had been terminated for unacceptable behaviour.
"After issuing a warning yesterday, we have no hesitation in taking this action after becoming aware of new information today."
Gray's sacking was followed by an announcement from Professional Game Match Officials that they had withdrawn 25-year-old Massey from tonight's npower League Two match between Crewe and Bradford, where she had been due to run the line.
PGMO general manager Mike Riley said: "PGMO and Sian believe that, with any football match, the focus should not be on the officials but on the players and the game itself.
"Sian is an excellent professional who has unwittingly found herself in the middle of a story that has nothing to do with her competence as a match official.
"Sian only wants to be notable for her performance as an assistant referee and is keen for things to get back to normal so she can return to officiating as soon as possible.
"Sian has the full backing of PGMO and we hope she continues her development, which has shown excellent progress so far.
"It is important for PGMO to have talented people from all backgrounds getting involved in refereeing.
"The more people are attracted to the game, the better standards will be, and the more everyone can talk about the football."
Reporter Burton was earlier disciplined for describing Massey as a "bit of a looker" in an off-air, but nevertheless recorded, conversation with Gray prior to Saturday's Premier League game between Wolves and Liverpool.
Gray responded by saying: "I definitely wouldn't."
Burton was stood down from tomorrow night's Carling Cup semi-final second leg between Birmingham and West Ham.
That followed Sky Sports' decision to remove Gray and Keys from duty for last night's Premier League game between Bolton and Chelsea after both were recorded off-air questioning Massey's appointment for the Wolves game, apparently agreeing that women "don't understand the offside rule".
Gray, whose comments towards Jackson were also off-air, loses a job that reportedly earned him £1.7million per year.
Sky Sports declined to comment this morning on when or if Keys would be back fronting their football coverage.
The broadcaster insisted Keys had not been sacked for his disparaging remarks but was unable to confirm exactly when he - who like Gray has been an employee for two decades - would return to the air.
His next presenting opportunity would appear to be the Premier League match between West Brom and Wigan on Tuesday week.
Keys telephoned Massey on Sunday night to apologise for his remarks and although Gray did not call Massey, Sky Sports insisted today Keys had done so on behalf of them both.
A spokesman told Press Association Sport Massey had accepted Keys' apology, adding: "It wasn't a case of Andy not apologising.
"It doesn't really make sense for both of them to call."
It was not known this afternoon whether Burton would elect to - or be forced to - make a similar apology and neither was it known whether Keys has apologised to West Ham vice-chairman Karren Brady, who he also disparaged for suggesting sexism was rife in the game.
Brady said Keys and Gray's comments about Massey made her "blood boil" and she was joined by a number of anti-discrimination groups in condemning the pair's remarks.
The decision to stand the duo down from last night's broadcast saw Sky dispense with their usual Isleworth-based studio operation, with a new team sent at the last minute to Bolton.
The programme was presented by David Jones instead of Keys, while Jamie Redknapp and Eidur Gudjohnsen were drafted in as pundits.
Meanwhile, it has emerged that the Football Association switchboard was inundated with calls yesterday from women enquiring about becoming officials.
Video: The footage believed to be behind Gray's dismissal...
Sky Sports statement in full:
The decision, which is effective immediately, was made in response to new evidence of unacceptable and offensive behaviour.
The new evidence, relating to an off-air incident that took place in December 2010, came to light after Andy Gray had already been subjected to disciplinary action for his comments of 22 January 2011.
Barney Francis, Managing Director of Sky Sports, commented: "Andy Gray's contract has been terminated for unacceptable behaviour. After issuing a warning yesterday, we have no hesitation in taking this action after becoming aware of new information today."
More to follow...