title: Human Ro60 (SSA2) genomic organization and sequence alterations, examined in cutaneous lupus erythematosus.
abstract: BACKGROUND
  The Ro 60 kDa protein (Ro60 or SSA2) is the major component of the Ro ribonucleoprotein (Ro RNP) complex, to which an immune response is a specific feature of several autoimmune diseases. The genomic organization and any sequence variation within the DNA encoding Ro60 are unknown.
  
  
  OBJECTIVE
  To characterize the Ro60 gene structure and to assess whether any sequence alterations might be associated with serum anti-Ro antibody in subacute cutaneous lupus erythematosus (SCLE), thus potentially providing new insight into disease pathogenesis.
  
  
  METHODS
  The cDNA sequence for Ro60 was obtained from the NCBI database and used for a BLAST search for a clone containing the entire genomic sequence. The intron-exon borders were confirmed by designing intronic primer pairs to flank each exon, which were then used to amplify genomic DNA for automated sequencing from 36 caucasian patients with SCLE (anti-Ro positive) and 49 with discoid LE (DLE, anti-Ro negative), in addition to 36 healthy caucasian controls.
  
  
  RESULTS
  Heteroduplex analysis of polymerase chain reaction (PCR) products from patients and controls spanning all Ro60 exons (1-8) revealed a common bandshift in the PCR products spanning exon 7. Sequencing of the corresponding PCR products demonstrated an A > G substitution at nucleotide position 1318-7, within the consensus acceptor splice site of exon 7 (GenBank XM001901). The allele frequencies were major allele A (0.71) and minor allele G (0.29) in 72 control chromosomes, with no significant differences found between SCLE patients, DLE patients and controls.
  
  
  CONCLUSIONS
  The genomic organization of the DNA encoding the Ro60 protein is described, including a common polymorphism within the consensus acceptor splice site of exon 7. Our delineation of a strategy for the genomic amplification of Ro60 forms a basis for further examination of the pathological functions of the Ro RNP in autoimmune disease.
authors.affiliation: Department of Photobiology, Guy's, King's and St Thomas' School of Medicine, St Thomas' Hospital, London SE1 7EH, UK. thomas.millard@kcl.ac.uk
authors.forename: T P
authors.initials: TP
authors.lastname: Millard
authors.forename: G H S
authors.initials: GH
authors.lastname: Ashton
authors.forename: E
authors.initials: E
authors.lastname: Kondeatis
authors.forename: R W
authors.initials: RW
authors.lastname: Vaughan
authors.forename: G R V
authors.initials: GR
authors.lastname: Hughes
authors.forename: M A
authors.initials: MA
authors.lastname: Khamashta
authors.forename: J L M
authors.initials: JL
authors.lastname: Hawk
authors.forename: J M
authors.initials: JM
authors.lastname: McGregor
authors.forename: J A
authors.initials: JA
authors.lastname: McGrath
mesh_terms: D000974:Antibodies, Antinuclear / Q000097:blood; D001324:Autoantigens / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D005787:Gene Frequency; D023281:Genomics; D020180:Heteroduplex Analysis; D006801:Humans; D008178:Lupus Erythematosus, Cutaneous / Q000235:genetics* / Q000276:immunology; D008179:Lupus Erythematosus, Discoid / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D020641:Polymorphism, Single Nucleotide; D020733:RNA, Small Cytoplasmic; D012261:Ribonucleoproteins / Q000235:genetics* / Q000276:immunology; D017422:Sequence Analysis, DNA; D000099207:SS-A Antigen
chemical_list: D000974:Antibodies, Antinuclear; D001324:Autoantigens; D020733:RNA, Small Cytoplasmic; D012261:Ribonucleoproteins; C116006:RO60 protein, human; D000099207:SS-A Antigen