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Former cabinet minister David Davis has joined calls for Boris Johnson to stand down, telling the prime minister: "In the name of God, go."
He said the PM had failed to take responsibility for his actions over lockdown parties in Downing Street.
It came after backbench Tory MP Christian Wakeford defected to Labour, minutes before Prime Minister's Questions began.
The PM batted away repeated calls to quit during the stormy Commons session.
In a letter to Mr Johnson, Bury South MP Mr Wakeford said: "You and the Conservative Party as a whole have shown themselves incapable of offering the leadership and government this country deserves."
Sir Keir Starmer welcomed the MP to his new party and repeated his call for Mr Johnson to quit, saying his "absurd and unreliable defences" of No 10 parties were unravelling.
But the most dramatic intervention came towards the end of the session, when veteran Tory MP David Davis told Mr Johnson he had spent weeks defending him from "angry constituents", including by reminding them of the "successes of Brexit".
He added: "I expect my leaders to shoulder the responsibility for the actions they take. Yesterday he did the opposite of that. So, I will remind him of a quotation which may be familiar to his ear: Leopold Amery to Neville Chamberlain.
"You have sat too long here for any good you have been doing. In the name of God, go."
The PM said he "didn't know what [Mr Davis] is talking about", but insisted he took "full responsibility for everything done in this government and throughout the pandemic".
Speaking afterwards to the BBC's Carolyn Quinn, Mr Davis said he had supported Mr Johnson until his interview on Tuesday, when he said he had not been told a party in the Downing Street garden risked breaking Covid rules.
"Yesterday's interview was an attempt to escape responsibility, not to shoulder it. And that's a test of leadership for me."
But he confirmed he hasn't put in a letter of no confidence to the chairman of the backbench 1922 committee, Sir Graham Brady, who organises Tory leadership contests.