Protein heterogeneity in the human Ro/SSA ribonucleoproteins. The 52- and 60-kD Ro/SSA autoantigens are encoded by separate genes.
Two cDNA clones encoding the 52-kD form of a protein present in human Ro/SSA ribonucleoprotein complexes were cloned from a lambda gt11 human thymocyte cDNA library. These clones reacted with lupus patient sera which had anti-52-kD Ro/SSA antibodies, and with affinity-purified anti-52-kD Ro/SSA antibodies. Moreover, affinity-purified antibodies isolated from isopropyl-beta-D-thiogalactopyranoside-induced proteins of these clones reacted only with the 52-kD protein of lymphocytes in Western blots and precipitated Ro/SSA hY RNAs, confirming that the clones encode a 52-kD Ro/SSA antigen. The cDNA contains a single open reading frame of 1,425 nucleotides and encodes a predicted 475-amino acid polypeptide with a molecular mass of 54,108 D. This protein appears unique in that both a zinc finger and leucine zipper motif are present on this protein. Surprisingly, no homology was found between the 52-kD Ro/SSA gene or protein and three published 60-kD Ro/SSA sequences. However, significant similarity of the 52-kD Ro/SSA was detected with human rfp and mouse rpt-1. These three proteins each contain similar zinc finger motifs in approximately their first 145 amino acid residues. The cDNA and the protein expressed therefrom are useful in the analysis of the structural and functional properties of this autoantigen.
Itoh
K
K
Arthritis and Immunology Program, Oklahoma Medical Research Foundation, Oklahoma City 73104.
Itoh
Y
Y
Frank
M B
MB
D000595:Amino Acid Sequence; D000818:Animals; D000974:Antibodies, Antinuclear / Q000032:analysis; D001324:Autoantigens / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D004247:DNA / Q000032:analysis / Q000302:isolation & purification; D006801:Humans; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D020733:RNA, Small Cytoplasmic; D012261:Ribonucleoproteins / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D012689:Sequence Homology, Nucleic Acid; D000099207:SS-A Antigen
D000974:Antibodies, Antinuclear; D001324:Autoantigens; D004247:DNA; D007544:Isopropyl Thiogalactoside; D020733:RNA, Small Cytoplasmic; D012261:Ribonucleoproteins; D000099207:SS-A Antigen; C116006:RO60 protein, human; C035356:SS-A antibodies