Annunziata, an opposition supporter, challenged Berlusconi on the RAI 3 programme to explain why to people outside Italy he was known only for his support of U.S. President George W. Bush in the Iraq war and for the conflict between his business and political interests.
Berlusconi, visibly irritated, refused to discuss his relationship with Bush further.
Cracks have started to show in his usually buoyant television style, before he faces opposition leader Romano Prodi on Tuesday in a U.S.-style television showdown on RAI.
"You are not used to taking journalists' questions, this is my show, I'll decide what questions I ask," snapped Annunziata, who quit her RAI job two years ago claiming it had become just a "mail box" for requests from the government.
However, Berlusconi still managed to get in the last word.
"And they say I'm the one who controls RAI!" he could be heard shouting through his still hooked-up microphone after he had left the set.
What a rat!!