The former Manchester City striker has paid the price for a poor start to the season that has seen the 2013 FA Cup winners drop into the relegation zone.
Wigan were expected to be among the contenders for promotion after being beaten by QPR in the play-off semi-finals last season, but have won only three of their first 17 games in the second tier this term.
A 3-1 derby defeat against Bolton Wanderers last Friday ensured Wigan have secured just one victory in their last 12 league games and chairman Dave Whelan has lost patience with Rosler, who he recruited from Brentford last December.
The Wigan chairman said in a statement on the club's official website: "I have thought long and hard about this, I have thought about little else for days in fact.
"Initially my thoughts were that Uwe should be given more time. He is a good manager and I like him immensely as a person, he is honest, passionate and clearly works day and night to try and earn success for the team.
"Following the incredible achievements of last season, I felt it was the right thing to do that he should be given more time. However, the situation we find ourselves in at the moment, and the run of results in recent months, really has been alarming.
"Something is clearly not working. For whatever reason, the team is not gelling, despite substantial financial backing in the summer and the dressing room being blessed with a huge amount of quality.
"Therefore, I have now come to the reluctant conclusion that for the long term good of the club, we need a change.
"Uwe is a good man and a good manager and he will bounce back, I am sure of that. We thank him for his efforts with us, I could not have wished to appoint a more committed and hard-working manager and we sincerely wish him all the best in his future career.
"But now the search begins immediately for a replacement."
Assistant manager Graham Barrow will take charge of the first team while Whelan goes about finding a successor to Rosler.
