Trams between Tinsley and Meadowhall Interchange will be cancelled for two weeks later this month while roadworks are carried out.
They will not run between January 13 and 26, but replacement buses will run between the two tram stops at the same frequency as the trams would have operated – in a bid to keep disruption for passengers to a minimum.
Supertram tickets will be valid on the bus services.
The cancellations are to allow preparatory work to be carried out ahead of the construction of a new section of road under Tinsley Viaduct.
The road will eventually allow for quicker journeys between Sheffield and Rotherham, avoiding junction 34 of the M1.
The new road is due to open in 2015, following the approval of funding from the Department for Transport in October.
The replacement buses operating during the roadworks will run from the bus stop beside the Tinsley stop into stand D2 at Meadowhall Interchange.
Services are due to revert to normal timetables on Monday, January 27.