Poor posture has been suggested as one of the main factors contributing to the high prevalence of neck pain in video display unit (VDU) users, but no clear association between pain and any particular resting neck posture has been found. Postural awareness of the neck, as indicated by the repositioning accuracy, may therefore be an appropriate measure and potentially useful assessment tool. The objective of this study is to examine whether posture and fatigue affect the head repositioning ability in typical VDU usage. A group of 20 healthy participants reproduced a normal comfortable posture for forward, upright and backward chair back inclinations in random order both before and after fatigue of the upper trapezius muscles. Ten repetitions of the posture were recorded for 2 s each, and the angular and translational deviations from the original head position were measured with regard to the external environment (head in space repositioning) and with regard to the trunk (head on trunk repositioning). Analysis by repeated measures ANOVA showed significant effects and interactions of fatigue and chair back inclination on the repositioning errors in the sagittal plane, which typically showed systematic trends towards certain postures rather than random errors around a mean position. While further work is required to examine the ergonomic impact of impaired repositioning ability, head repositioning is sensitive to ergonomic factors such as seating configuration and fatigue, and may therefore be a useful tool for evaluation of static working postures.