Expression and functional conservation of the human La (SS-B) nuclear autoantigen in murine cell lines.
We have studied the cellular expression and functional conservation of the human La autoantigen across species by introducing genes encoding human La antigen into cultured murine cell lines. In transfected murine fibroblasts and lymphoid cell lines human La was expressed as a predominantly nuclear antigen, with a typical pattern of nuclear speckling. Radiolabeled human La was of the predicted molecular mass (48 kDa) when expressed in murine cells and was associated intracellularly with murine 60 kDa Ro antigen. Under certain conditions of cell lysis the association between human La and murine Ro was disrupted, whereas human La and human Ro complexes remained intact, suggesting the possibility of species-specific protein interaction between La/Ro antigens. Expression of human La from the genomic gene construct was associated in some transfectants with the presence of additional high molecular weight bands reactive with anti-La monoclonal antibodies. Cell surface La determinants were not detected on any of the transfected cells. Human La protein associated with many of the same radiolabeled RNAs as the murine La antigen in transfected murine cells, indicating intact RNA-binding function of La protein across species. The findings indicate that despite 23% non-identity within the primary structure of human and murine La, the human molecule is functionally highly conserved across species.
Keech
C L
CL
Department of Clinical Immunology, Flinders Medical Centre, Bedford Park, South Australia.
Gordon
T P
TP
Reynolds
P
P
McCluskey
J
J
D000818:Animals; D001324:Autoantigens / Q000032:analysis / Q000096:biosynthesis / Q000502:physiology*; D002460:Cell Line; D003001:Cloning, Molecular; D005347:Fibroblasts; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence; D009710:Nucleotide Mapping; D020733:RNA, Small Cytoplasmic; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins / Q000032:analysis / Q000096:biosynthesis / Q000502:physiology*; D014162:Transfection; D000097585:SS-B Antigen; D000099207:SS-A Antigen
D001324:Autoantigens; D020733:RNA, Small Cytoplasmic; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; D000097585:SS-B Antigen; D000099207:SS-A Antigen; C116006:RO60 protein, human