Luzhkov, 74, a pillar of the ruling United Russia party, had ruled the capital since 1992, but angered Medvedev by suggesting the country needed a stronger and more decisive leader.
Medvedev, on a state visit to China, issued a decree stripping Luzhkov of his post. Russian news agencies said Luzhkov learned he had been sacked from a television report.
"As president of Russia I lost my trust in Yuri Mikhailovich Luzhkov as the mayor of Moscow," Medvedev told journalists. "I will decide who will lead Moscow."
Putin offered support for Medvedev's decision, suggesting the mayor had overstepped the mark by challenging the Kremlin.
Luzhkov, famous for his flat working man's caps, billionaire second wife and blunt manner, used a $37-billion budget to keep pensions and public services high, remaining relatively popular despite corruption allegations, which he always denied.
Reuters
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