Tymoshenko responded by saying Lutsenko would continue to head the ministry as its first deputy minister, Ukrainian news agencies reported.
"Today at a government meeting, Lutsenko will be named the first deputy, the acting head and he will head the interior ministry," she was quoted as saying by local agencies.
Yanukovich aides said the loss of Lutsenko from his role as interior minister would deprive Tymoshenko of chances of rigging the vote and doom her hopes of victory.
"Parliament has brought to a stop the inglorious career of Lutsenko," Mykola Azarov, a top Yanukovich aide, said.
"In a few days time, the Ukrainian people will bring to a stop the inglorious career of Tymoshenko."
Volodymyr Fesenko, the director of Penta, a Ukrainian think tank, said the decision "means the Regions party has succeeded in weakening its rival in a significant part of the election process."
The opposition in parliament has tried to sack Lutsenko several times, most recently last year after German police said they had detained him at a Frankfurt airport for drunk and disorderly behaviour.