Previously, we compared signal-mediated nuclear transport in proliferating and quiescent BALB/c 3T3 cells and found that both the relative rate of nuclear uptake and the functional size of the transport channels were significantly greater in proliferating cells. In this study, the possible causes of these permeability differences were investigated. To determine if the decrease in transport capacity in quiescent cells was due to a reduction in the availability of soluble cytoplasmic factors (i.e., ATP or receptors for nuclear location sequences), or changes in the properties of the pores themselves, proliferating and quiescent cells were fused, and nuclear import of nucleoplasmin-coated gold (NP-gold) particles was assayed in the heterokaryons 50-60 min later. Significant differences in nuclear uptake were maintained following fusion, even though the two nuclei shared a common cytoplasm, consistent with the view that permeability is regulated at the level of the pores. Cell shape also influenced signal-mediated nuclear import. This was demonstrated by studying transport in rounded and flattened cells attached to different-size palladium domains that were deposited on a nonadhesive substrate. Based on analysis of the nuclear uptake rates of large (110-270 A in diameter) and small (50-80 A in diameter) coated gold particles, it was determined that the functional size of the pores was significantly greater in flattened cells. The effect of growth factors on recovery of nuclear transport capacity following serum depletion was also analyzed. Partial recovery was achieved by treating cells with physiological concentrations of EGF, IGF-1, or PDGF; however, complete recovery required both EGF and IGF-1.