Metrolink trams will not run on the Eccles line during the February half-term.
The line is being fully closed between February 19 and 27 for "essential maintenance" of the tracks.
Bosses say the school holidays have been chosen in order to 'minimise' disruption.
Further partial closures which will also be needed are to take place during the Easter and summer school breaks, Transport for Greater Manchester (TfGM) said.
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The Eccles line opened in 1999, with the spur to MediaCity opening in 2010, and they have seen millions of journeys during that time.
However, some of the track is now 22 years old and needs replacing.
Bosses say "it’s a big job, but it has to be done".
During the February closure, those who are able to are being urged to use the Trafford Park Line
However, replacement bus services will run between Eccles and Piccadilly Station calling at all of the line's stops.
Services on the Ashton line, which normally run through to Eccles, will instead run through to Bury.
Danny Vaughan, Head of Metrolink at Transport for Greater Manchester, said: “In the past 10 years alone the Eccles line has supported 40 million journeys – providing a real alternative to using the car, helping people get around the region, reducing congestion and helping to cut air pollution.
“The Eccles Line, and the MediaCity spur, which was opened in 2010, have supported the local economy and been instrumental in the regeneration of the Quays area.
"It is therefore very important that we do these works now in order to help the area continue to grow and prosper.
“We know it’s going to cause some inconvenience for passengers so we are limiting the work to less busy times.
"Many passengers will be able to catch trams on the Trafford Park Line, and we will provide good alternatives to those who can’t.”
The nearly four-mile-long Eccles line first opened as far as the Broadway stop in December 1999 as part of the phase two expansion of the network and was opened by then Prime Minister Tony Blair.
The service was then extended to Eccles Interchange in July 2000 and was officially opened by Princess Anne in July 2001.
A short spur to MediaCity opened in 2010.
