Alpha-actinins are actin-binding proteins of non-muscle cells, which can participate in the regulation of transcription factor activity. We describe the distribution of α-actinin-1 and −4 depending on different actin cytoskeleton formed as a result of cell adhesion to extracellular matrix proteins, such as fibronectin and laminin 2/4. Immunofluorescent studies show a difference in the distribution of α-actinin and −4. Both isoforms localise along stress-fibres, but α-actinin-1 localises in the perinuclear region more abundantly than α-actinin-4. Western blot analysis demonstrated existence of truncated forms of both isoforms. Truncated α-actinin-1 appears in cells spread on fibronectin or laminin. Cell spreading also correlated with more tight association of α-actinin-4 with chromatin. Basing on our previous finding of an interaction of α-actinin-4 with p65 subunit of the NF-κB, we checked the possible influence of immobilised ligands on its redistribution in nuclear complexes containing p65. α-Actinin-4 seems to be present in some but not all nuclear complexes containing p65. Immobilised ligands may affect the interaction of α-actinin-4/p65 complexes with chromatin. The data suggest that adhesion to extra-cellular matrix may interfere in cellular reactions mediated by α-actinin-1 and −4.