When Perosi died in 1956, Pope Pius XII gave Bartolucci the position of permanent director of the Pontifical Sistine Chapel Choir.

[5] Among those most against the decision, motivated by Papal Master of Ceremonies Archbishop Piero Marini, was Cardinal Joseph Ratzinger,[6] who, after he became Pope Benedict XVI, recalled Bartolucci to direct a concert in the Sistine Chapel on 24 June 2006, in which he offered music from the repertoire of sacred polyphony of Giovanni Pierluigi da Palestrina alongside his own compositions including the motet Oremus pro Pontifice Nostro Benedicto ("Let us pray for our Pontiff Benedict"), dedicated to the Pope.

In the forty years of Bartolucci's leadership, the choir balanced the obligation of papal liturgies with tours in various countries throughout the world, including Austria, France, Belgium, the Philippines, Australia, the United States, Turkey, Poland and Japan.

[9] The body of his work already published fills more than forty volumes and includes Masses, motets, madrigals, hymns, symphonic, organ, and chamber music, and above all a series of oratorios for soloists, chorus, and orchestra.

Characteristic of Bartolucci's aesthetic conception is a respect for tradition, whose base lies in "a considerable severity of song and a certain limpid and solid polyphony", as he describes in the preface to his First Book of Motets.

Because he was over the age of 80, under the terms of Pope Paul VI's 1970 motu proprio Ingravescentem aetatem, he was never eligible to vote in any future Papal conclave.

His funeral Mass took place at St. Peter's Basilica on 13 November, and was led by Angelo Sodano, Dean of the College of Cardinals, with Pope Francis performing the final commendation.