A new concept in the interior design of autonomous vehicles is rotatable or swivelling seats that allow people sitting in the front row to rotate their seats and face backwards. In the current study, we used a take-over request task conducted in a fixed-based driving simulator to compare two conditions, driver front-facing and rear-facing. Thirty-six adult drivers participated in the experiment using a within-subject design with take-over time budget varied. Take-over reaction time, remaining action time, crash, situation awareness and trust in automation were measured. Repeated measures ANOVA and Generalized Linear Mixed Model were conducted to analyze the results. The results showed that the rear-facing configuration led to longer take-over reaction time (on average 1.56 s longer than front-facing, p < 0.001), but it caused drivers to intervene faster after they turned back their seat in comparison to the traditional front-facing configuration. Situation awareness in both front-facing and rear-facing autonomous driving conditions were significantly lower (p < 0.001) than the manual driving condition, but there was no significant difference between the two autonomous driving conditions (p = 1.000). There was no significant difference of automation trust between front-facing and rear-facing conditions (p = 0.166). The current study showed that in a fixed-based simulator representing a conditionally autonomous car, when using the rear-facing driver seat configuration (where participants rotated the seat by themselves), participants had longer take-over reaction time overall due to physical turning, but they intervened faster after they turned back their seat for take-over response in comparison to the traditional front-facing seat configuration. This behavioral change might be at the cost of reduced take-over response quality. Crash rate was not significantly different in the current laboratory study (overall the average rate of crash was 11%). A limitation of the current study is that the driving simulator does not support other measures of take-over request (TOR) quality such as minimal time to collision and maximum magnitude of acceleration. Based on the current study, future studies are needed to further examine the effect of rotatable seat configurations with more detailed analysis of both TOR speed and quality measures as well as in real world driving conditions for better understanding of their safety implications.