The relative biological effectiveness (RBE) of neutrons and other types of densely ionizing radiation appears to be close to 1.0 for the induction of strand breaks, but considerably higher RBEs have been found for cellular end points such as colony-forming ability. This may be due to differences in the processing of strand breaks or to the involvement of other lesions whose yields are more dependent on radiation quality. Because cell cycle delays may be of great importance in the processing of DNA damage, we determined the RBE for disturbances of the G1 phase in four different cell types (Be11 melanoma, 4197 squamous cell carcinoma, EA14 glioma, GM6419 fibroblasts) and compared them with the RBE for cell inactivation. The method we used to determine the progress from G1 into S was as follows: Cells were serum-deprived for a number of days and then stimulated to grow with culture medium containing normal amounts of serum. Immediately before the change of medium, cells were exposed to graded doses of either 240 kV X rays or 6 MeV neutrons. At different times afterward, cells were labeled with BrdU and the numbers of active S-phase cells were assessed using two-parameter flow cytometry. For all four cell types, cells started to progress from G1 into S after a few hours. Radiation suppressed this process in all cases, but there were some interesting differences. For Be11 and 4197 cells, the most obvious effect was a delay in G1; the labeling index increased a few hours later in irradiated samples than in controls, and there was no significant effect on the maximum labeling index. For EA14 and GM6419 cells, although smaller doses were used because of greater radiosensitivity, a delay of the entry into S phase was again noticeable, but the most significant effect was a reduction in the maximum percentage of active S-phase cells after stimulation, indicating a permanent or long-term arrest in G1. The RBE for the G1 delay was the same for all four cell types, about 2.8, while the RBE for the G1 arrest varied between 3.2 for the most resistant Be11 cells and 1.7 for the most sensitive GM6419 cells. This trend was similar to that observed for the RBE for cell inactivation. If, as described above, the same number of strand breaks per dose is induced by neutrons and by X rays, the signal transduction cascade translates them into a greater G1 delay in the case of higher LET. This appears to be independent of repair capacity, because it is similar in all cell types we investigated. We therefore assume that a higher lesion density or the presence of other types of lesions is important for this relatively early effect. A G1 arrest, however, is more closely related to the later events leading to cell inactivation, where strand break repair does play a major role, influencing X-ray sensitivity more strongly than sensitivity to neutrons because of a lower repairability of lesions induced by higher-LET radiation.