Protein 4.1 is a component of the nuclear matrix of mammalian cells

Abstract

Protein 4.1 is a major component of the erythrocyte membrane skeleton that promotes the interaction of spectrin with actin and links the resulting complex network to integral membrane proteins. Here we analyse the distribution of different 4.1 proteins within the nucleus of mammalian cells. Nuclear matrices have been prepared from Madin-Darby canine kidney (MDCK) and HeLa cells and protein fractions isolated at each step of the purifications have been analysed by immunoblotting using characterized polyclonal antibodies against protein 4.1. Two 4.1 polypeptides of M(r) approximately 135,000 and 175,000 are extracted after DNase I digestion and 0.25 M ammonium sulphate treatments, suggesting that they may be associated with chromatin. Interestingly, nuclear matrices isolated after DNase I digestion and sequential treatments with increasing ionic strength contain a third 4.1 polypeptide of M(r) approximately 75,000 (4.1p75), suggesting that it is a component of the nuclear matrix. Immunoblot analyses of nuclear matrices isolated from different cell types and species indicate that 4.1p75 is a common element of the nuclear matrix of mammalian cells. Moreover, 4.1p75 distributes to typical nuclear speckles which are enriched with the spliceosome assembly factor SC35, as revealed by double-label immunofluorescence analyses. Protein 4.1p75 might be an anchoring element of the nucleoskeleton, playing a role similar to that described for the erythroid protein 4.1 in red blood cells.