A senior Tory has accused the Government of "blackmail" of Conservative rebels and said they should report it to the police.
William Wragg, chair of the Commons Public Administration Committee, said Tory colleagues had been subject to intimidation by the party whips over their perceived opposition to the Prime Minister.
No10 staff, special advisers and government whips also encouraged embarrassing stories being released to journalists if they did not support the PM, he claimed.
Mr Johnson was accused of "acting like a mafia boss" as the extraordinary claims sparked outrage.
But the PM, on a visit to Taunton, said: "I've seen no evidence to support any of those allegations.
"What I am focused on is what we're doing to deal with the number one priority of the British people, which is coming through Covid."
His future is hanging in the balance after the Downing Street party saga, with his allies mounting a desperate bid to woo mutinous Tories openly plotting against him.
At least six Conservative MPs have confirmed that they have submitted letters of no confidence in the PM, including Mr Wragg who broke cover last week to call for him to quit.
In a statement in Parliament, Mr Wragg said Tory rebels had been threatened with losing money for their constituencies which comes from the taxpayer.
Mr Wragg said: "In recent days, a number of members of Parliament have faced pressures and intimidation from members of the government because of their declared or assumed desire for a vote of confidence in the party leadership of the Prime Minister."
He said the whips were responsible for pushing forward Government business but added: "It is not their function to breach the ministerial code in threatening to withdraw investments from members of parliament's constituencies which are funded from the public purse."
Mr Wragg added: "Intimidation of a member of Parliament is a serious matter. Moreover, the reports of which I am aware would seem to constitute blackmail.
"As such it would be my general advice to colleagues to report these matters to the Speaker of the House of Commons and the Commissioner of the Metropolitan Police."
His claims were backed up by Tory defector Christian Wakeford - who sensationally claimed he was threatened he would not get a new high school in his constituency if he didn't vote with the Government.
He told BBC North West: "I was threatened that I would not get the school for Radcliffe if I did not vote in one particular way.
"This is a town that's not had a high school for the best part of 10 years.
"How would you feel when holding back regeneration of a town for a vote?
"It didn't sit comfortably and that was really starting to question my place where I was and ultimately to where I am now."
Cabinet Office Minister Stephen Barclay said he would convey the statement to the Government.
Labour's Deputy Leader Angela Rayner said: "These are grave and shocking accusations of bullying, blackmail, and misuse of public money and must be investigated thoroughly.
"The idea that areas of our country will be starved of funding because their MPs don't fall into line to prop up this failing Prime Minister is disgusting.
“On the day Labour is setting out plans to deal with the cost of living crisis affecting the whole country, the Tory party continues to descend further into chaos of its own making. The country deserves better than this out of touch, out of control, out of ideas government.”
Lib Dem leader Sir Ed Davey accused the PM of "acting like a mafia boss".
"It is simply remarkable that a Prime Minister could countenance scrapping local projects and funding if MPs refuse to back him," he said.
"All Boris Johnson cares about is saving his own skin. He’s acting more like a mafia boss than a Prime Minister.
"These latest allegations of criminal behaviour are yet more proof that a civil servant investigation into Partygate is utterly inadequate. The Metropolitan Police must investigate.
"Number 10 is now in full scorched earth mode. Every minute the Tories allow him to stay in power damages our country."
A No 10 spokesperson said: "We are not aware of any evidence to support what are clearly serious allegations.
"If there is any evidence to support these claims we would look at it very carefully."
But Tory MP Michael Fabricant corroborated Mr Wragg’s bombshell claims - in the same sentence as trying to dismiss them.
He tweeted: “If I reported every time I had been threatened by a Whip or if a Whip reported every time I had threatened them, the police wouldn’t have any time to conduct any other police work! What nonsense from WW.”
Asked if Boris Johnson was behaving like a mafia boss, his spokesman said: “Well, er, obviously we’ve been clear again that there is no evidence to support the claims put forward.
"This is a matter for the whips’ office and not one for me to comment on.”
The PM’s spokesman added: “We’d expect everyone to act within any requisite rules and guidance.”
Asked if the PM condemned all forms of bullying and harassment the spokesman replied: “Yes.”
But asked if there would be an inquiry he replied: “Certainly not that I’m aware of.”
Asked “how does the Prime Minister sleep at night?”, his spokesman replied: “It’s not a question I’ve asked him”.
Commons Speaker Sir Lindsay Hoyle said it would be a "contempt" to obstruct MPs from doing their jobs by trying to "intimidate" them.
He expressed concern at the "serious allegations" made by Mr Wragg.
Sir Lindsay told MPs: "Those who work for them are not above the criminal law. The investigation of allegedly criminal conduct is a matter for the police and decisions about prosecution are for the CPS. It will be wrong of me to interfere with such matters.
"While the whipping system is long-established, it is of course a contempt to obstruct members in the discharge of their duty or to attempt to intimidate a member in their parliamentary conduct by threats."
It comes as Labour was working to win more Tory defectors in the aftermath of Christian Wakeford’s bombshell decision to join Keir Starmer's party.
A source told the Mirror the "door is always open", but added: “I’m not expecting an imminent second one - but that doesn’t mean there won’t be one.
“We’ll have to see what happens over the next few weeks.”
