The Cherry Street level crossing will be closed between 8pm and 6am each night from Thursday 17 December to Monday 21 December.
Drivers will be detoured via Derrimut Road, Princes Highway and Werribee Street.
Access to Bunnings and Woolworths on Cherry Street will remain during the works.
Following a recent signaling fault on the Werribee line, the Level Crossing Removal Authority need to complete additional signal testing at the boom gates and within the rail corridor.
The works are being completed overnight to reduce impacts to drivers during peak travel times.