On the site of the modern Holy Assumption Cathedral, which was the center of the Poltava fortress, there previously stood a wooden Assumption church known as the "town cathedral." The last mentions of it date back to 1695. In 1776, the wooden church was handed over to the community of the village of Ivashki in the Poltava district. Its status as a cathedral church is evidenced by an inscription in an ancient akathist book. In 1748, at the initiative of Colonel Andriy Horlenko and other representatives of the Cossack elite and the city community, a decision was made to build a stone church. On November 8, 1748, a contract was signed with the famous architect Stefan Stabansky. The work began in 1751 and continued until 1770.

This was the first stone cathedral in Poltava, measuring 32 meters in length and 17 meters in width. The building was two-storied in the style of a Byzantine basilica with decorative pediments. Around 1780, the church was reconstructed: two domes were added over the altar part, making it five-domed. The second floor was equipped with a choir, for which a staircase was built, and the vestry was reconstructed. The floor was originally made of pottery bricks but was later replaced with cast iron.

In 1899-1900, the cathedral was expanded: a side chapel was added, making it brighter and more spacious, increasing the area by almost 170 sq.m. Two small towers and an arched ceiling appeared above the entrance, two columns were dismantled, and the choir was moved to the western wall. In 1847, after the episcopal see was transferred from Pereyaslav to Poltava, the "town cathedral" became a cathedral. The cathedral was surrounded by a brick fence with wrought iron grilles, replacing the wooden one in 1827. Inside, there was a four-tiered carved iconostasis "in the French style." The cathedral had five altars: the main one in the name of the Assumption of the Blessed Virgin Mary; to the right, the apostles Peter and Paul; to the left, the prophet Elijah, consecrated on September 17, 1774; in the choir on the right, St. Archangel Michael, consecrated on December 30, 1776; on the left, St. Great Martyr Barbara, consecrated on June 10, 1777. The bell tower, 44 meters high, stood empty for a long time after the cathedral was destroyed in 1934. During the war from 1941 to 1946, the Church of John the Baptist, consecrated on February 27, 1787, operated in it.

After the war, the bell tower remained in ruins for a long time, later housing various workshops. In honor of the 1100th anniversary of Poltava, it was decided to restore the bell tower and create a city museum with an observation deck on the upper tier. Recently, it hosted an exhibition of amateur artists from the city. The bell tower remained until the early 1990s.

The decision to restore the cathedral was made in 1999 and completed in 2005.