Photolysis of molecular oxygen (O2) sustains the stratospheric ozone layer and is thereby protecting living organisms on Earth by absorbing harmful ultraviolet radiation. In the past, atmospheric O2 levels have not been constant, and their variations are thought to be responsible for the extinction of species due to the thinning of the ozone layer. Over the Phanerozoic Eon (last ~500 Mio years), the O2 volume mixing ratio ranged between 10% and 35% depending on the level of photosynthetic activity of plants and oceans. Previous estimates, however, showed ambiguous ozone (O3) responses to atmospheric O2 changes, such as monotonically positive or negative correlations, or displaying a maximum in the O3 column around a certain O2 level. Here, we assess the ozone layer sensitivity to atmospheric O2 varying between 5% and 40% with a state-of-the-art chemistry-climate model (CCM). Our findings show that the O3 layer thickness maximizes around the current mixing ratio of O2, 21%, under the contemporary boundary conditions. Lower or higher levels of O2 result globally in a reduction of total column O3 and, therefore, allow more harmful ultraviolet radiation (UV) to reach the surface. At low latitudes, the total column O3 is less sensitive to O2 changes, because of the known “self-healing” effect. In mid and high latitudes, however, there is an equator-to-pole variation between high and low O2 cases. Particularly, polar cap O3 is more sensitive to O2 with changes up to 20 DU even for small O2 perturbations. These variations are modulated by the radiative impact of O3 on stratospheric temperatures and on the strength of the Brewer-Dobson circulation (BDC), indicating chemistry-radiation-transport feedback. High O2 cases result in an acceleration of the BDC, and vice versa. This renders the relationship between O3 and molecular O2 to be non-linear on both global and regional scales, however still maximizing global mean total O3 column at contemporary O2 levels.