Judge Luis Aláez will grill five Adif officials over track and signal security on the line where the accident took place.
The current present, Gonzalo Ferre, and two former Adif presidents, Enrique Verdeguer and Antonio González Marín will also appear in court to answer questions.
Aláez has also said he will call up those directors on Adif's board who have served since the Santiago–Ourense line was opened in December 2011.
The move by Judge Aláez marks a stepping up in the investigation of what was Spain's deadliest rail accident since 1944.
The train came off the tracks on a sharp bend on the outskirts of the northwestern city of Santiago de Compostela.
The train's recorders have revealed it was travelling at 179 kilometres (111 miles) per hour when it derailed on a track with a speed limit of 80 kph, just after the driver finished a telephone call with a conductor.
Alaez has previously stated he wanted to question operators responsible for safety on the route that night to see whether they gave enough information to the driver on when to slow down.
Rail officials have said the track where the train crashed was not equipped with advanced braking systems that would slow the train down if the driver failed to do so.
Adif and the state company that runs the trains, Renfe, have both denied any technical failings and said correct procedures were followed.