Sun2 is a novel mammalian inner nuclear membrane protein.
Sun protein (Sun1 and Sun2) cDNAs were previously cloned based on the homology of their C-terminal regions (SUN (Sad1 and UNC) domain) with the Caenorhabditis elegans protein UNC-84 whose mutation disrupts nuclear migration/positioning. In this study, we raised an anti-Sun2 serum and identified Sun2 in mammalian cells. In HeLa cells, Sun2 displays a nuclear rim-like pattern typical for a nuclear envelope protein. The Sun2 antibody signal co-localizes with nuclear pore and INM markers signals. The rim-like pattern was also observed with the recombinant full-length Sun2 protein fused to either EGFP or V5 epitopes. In addition, we found that a recombinant truncated form of Sun2, extending from amino acids 26 to 339, is sufficient to specify the nuclear envelope localization. Biochemical analyses show that Sun2 is an 85-kDa protein that is partially insoluble in detergent with high salt concentration and in chaotropic agents. Furthermore, Sun2 is enriched in purified HeLa cell nuclei. Electron microscopy analysis shows that Sun2 localizes in the nuclear envelope with a sub-population present in small clusters. Additionally, we show that the SUN domain of Sun2 is localized to the periplasmic space between the inner and the outer nuclear membranes. From our data, we conclude that Sun2 is a new mammalian inner nuclear membrane protein. Because the SUN domain is conserved from fission yeast to mammals, we suggest that Sun2 belongs to a new class of nuclear envelope proteins with potential relevance to nuclear membrane function in the context of the involvement of its components in an increasing spectrum of human diseases.
Hodzic
Didier M
DM
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Yeater
David B
DB
Bengtsson
Luiza
L
Otto
Henning
H
Stahl
Philip D
PD
D000818:Animals; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D009685:Nuclear Envelope / Q000737:chemistry*; D009687:Nuclear Proteins / Q000737:chemistry* / Q000502:physiology; D010446:Peptide Fragments / Q000737:chemistry* / Q000502:physiology; D011817:Rabbits; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry*
D029742:Caenorhabditis elegans Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009687:Nuclear Proteins; D010446:Peptide Fragments; C093512:SAD1 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; C120560:Unc-84 protein, C elegans