The Blackwall Tunnel will be shut southbound for two weekends this summer.
Transport for London (TfL) has said that the closures will allow for a new road configuration to support the new Silvertown Tunnel.
Road resurfacing and safety barrier works are also expected to be carried out.
The works will mean no southbound traffic will flow through the tunnel on the weekends of July 20 to 22, and August 17 to 19.
Northbound traffic will not be affected by the closures and will be diverted via the southbound tunnel when required.
Plans for the Silvertown Tunnel, which is expected to open next year, were first proposed more than a decade ago.
It is expected that the new tunnel will help reduce traffic congestion and deliver faster journeys in east London, making travel times up to 20 minutes quicker.
There will also be a significant increase in bus services across the river and a proposed new cycle shuttle service through the tunnel for cyclists.
The Blackwall Tunnel weekend closures will start at 12.01am on the Saturday morning, with southbound traffic being restored by 5am on the Monday morning.
TfL has also said that it will take the opportunity to carry out works within and around the Blackwall tunnel, to help deliver future maintenance work more efficiently and ensure the tunnel can continue to safely operate.
Bus Route 108 will be temporarily split and terminate at Canning Town station and North Greenwich during the works.
Nick Fairholme from TfL said: “We are now in the final phase of works to complete the Silvertown Tunnel and these closures will help us to to start to get the tunnel ready to open.
“We appreciate that these planned weekend works will have an impact on some drivers and are doing everything we can to limit potential disruption - including ensuring a northbound route through the Blackwall Tunnel is maintained throughout.”
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