Boris Johnson, the London mayor, has become embroiled in a new row with David Cameron after government sources accused him of "overstepping the mark" by attacking plans for the £32bn north-south high-speed rail link.

The prime minister has identified the 250mph route linking London to Birmingham, Leeds and Manchester as a potential vote winner in the north but Johnson has joined mounting opposition.

In his submission to a public consultation on the High Speed Two (HS2) project, Johnson said aspects of the route were unacceptable while warning that it did not solve UK airport capacity problems.

He said: "These proposals need to give consideration to their impact on residents in London. There is a great case for investing in a high-speed rail network but that must also be supported by the infrastructure to handle the extra passengers."

Johnson is calling for a new tube link at Euston station, the project's proposed London terminus, amid fears that existing lines will be flooded with passengers.

A senior government source said Johnson, whose interventions on spending cuts and a ban on airport expansion have angered the cabinet, had "overstepped the line in his opposition to HS2."

It is understood that Philip Hammond, the transport secretary, has dismissed Johnson's request for more investment in London's transport, on top of funding for the £16bn Crossrail project and a multibillion-pound tube upgrade.

The source said: "Philip Hammond thinks high-speed rail is crucial for bridging the north-south divide. London did very well in the spending settlement and there is no case for the capital having even more major transport projects."

Rail companies have been asked to contribute £10,000 each to a pro-HS2 campaign amid fears the government will be outgunned by a well-funded opposition.

The "no" camp is strongest in the home counties, where residents are angry about the London-to-Birmingham phase, which is due to open in 2026.

The newly established Campaign for High Speed Rail accused Johnson of "hijacking" a national policy to secure more funding for London.

"The government is busy trying to address the north-south divide, so it's outrageous that Boris is trying to hijack this progress purely to secure more transport spending for London," said Professor David Begg, the group's director.

"London already receives more transport spending per resident than anywhere else in the country, and the HS2 scheme as it stands will benefit London hugely in terms of jobs and transport infrastructure."

Once the London-to-Birmingham route is completed, a Y-shaped extension opening in 2032 will take the line to Manchester and Leeds. The full route is forecast to produce a £44bn boost to the UK economy with the first phase generating an estimated 40,000 jobs, as well as freeing up capacity on the west coast rail line for more commuter and freight services.

However, opponents argue that the line will scar the landscape of Buckinghamshire, Warwickshire and Northamptonshire as well as the Chiltern Hills – all home to key Conservative constituencies.

The no campaign also believes that taxpayer funds would be better spent on a cheaper upgrade of the rail network.