The exact time of construction of the first main church of the Assumption Convent is unknown. Documents from the 18th century indicate that it was a two-story stone building. The upper floor housed the altar in honor of the Assumption of the Blessed Virgin Mary and a chapel in honor of St. Alexander of Svir, while on the lower floor, there was an altar in honor of the icon of the Mother of God "Burning Bush" and a chapel of St. Nicholas the Wonderworker. Over time, the building deteriorated, began to sink, and the first floor became flooded with water on rainy days and in spring. The church was dismantled, and in 1791-1792, a new one-story stone church was built with funds allocated by Catherine II, with the assistance of Tula's civil governor Andrei Ivanovich Lopukhin. The main altar was consecrated in honor of the Assumption of the Blessed Virgin Mary, the southern chapel was dedicated to the icon "Burning Bush," and the northern one to Alexander of Svir. In 1857, the church, already in a dilapidated state, was expanded, which only accelerated its destruction.

In 1899, the dismantling of this church began, and by 1902, a new two-story building was erected in its place. The lower warm church with one altar in the name of the Equal-to-the-Apostles Mary Magdalene and the Great Martyr Panteleimon was consecrated on October 24, 1902. The upper church, with the main altar of the Assumption, was consecrated on June 28, 1904, and the chapels of the upper church in honor of the icon "Burning Bush" and Alexander of Svir were consecrated on July 11, 1910, and July 3, 1911, respectively.

The Assumption Church was built in the Neo-Russian style. Its original design has been preserved, which is close to the constructed church but included somewhat different decorative elements. During construction, they abandoned Gothic decorative elements and pointed window frames on the drums. They also did not create a horizontal three-part division of the facades with windows, as this would have made the building three stories high. The walls of the upper church were painted in 1909. It is possible that the artists were associates of Viktor Vasnetsov, as the frescoes of the church are exact copies of his works in the Kiev Vladimir Cathedral. On the wall of the upper church, there is an inscription: "In 1909, through the efforts of the abbess Mother Magdalene, with the help of benefactors, this cathedral was adorned with wall paintings and ornaments." The walls of the cathedral are made of dark red brick, and the black domes are topped with crosses and crescents. The revered abbess Agnia, who was involved in the construction of the church, was buried in the cathedral.

With the coming to power of the Bolsheviks, the convent was abolished, and the cathedral was closed. In the 1930s, there were attempts to blow it up, but the cathedral proved to be very sturdy and withstood. Then the domes were removed, and a branch of the state archive was placed there. In the late 1980s, a reconstruction of the building was carried out, during which the domes were reinstalled and the foundation of the cathedral was strengthened.

On September 8, 2006, the cathedral was returned to the faithful, and on January 24, 2007, Archbishop Alexy of Tula and Belev consecrated the altar of the lower church, where the relics of St. John of Tula were transferred. On May 10, 2012, the second floor of the Holy Assumption Cathedral was solemnly consecrated, marking the completion of the restoration work on the church.