Driver sleepiness studies are often carried out with alert drivers during daytime and sleep-deprived drivers during night-time. This design results in a mixture of different factors (e.g. circadian effects, homeostatic effects, light conditions) that may confound the results. The aim of this study was to investigate the effect of light conditions on driver sleepiness. Thirty young male drivers (23.6 ± 1.7 years old) participated in a driving simulator experiment where they drove on a rural road. A 2 × 2 design was used with the conditions daylight versus darkness, and daytime (full sleep) versus night-time (sleep deprived). The results show that light condition had an independent effect on the sleepiness variables. The subjective sleepiness measured by Karolinska Sleepiness Scale was higher, lateral position more left-oriented, speed lower, electroencephalogram alpha and theta higher, and blink durations were longer during darkness. The number of line crossings did not change significantly with light condition. The day/night condition had profound effects on most sleepiness indicators while controlling for light condition. The number of line crossings was higher during night driving, Karolinska Sleepiness Scale was higher, blink durations were longer and speed was lower. There were no significant interactions, indicating that light conditions have an additive effect on sleepiness. In conclusion, Karolinska Sleepiness Scale and blink durations increase primarily with sleep deprivation, but also as an effect of darkness. Line crossings are mainly driven by the need for sleep and the reduced alertness at the circadian nadir. Lane position is, however, more determined by light conditions than by sleepiness.