title: Molecular cloning and primary structure of the human blood group RhD polypeptide.
abstract: The RH (rhesus) blood group locus from RhD-positive donors is composed of two homologous structural genes, one of which encodes the Cc and Ee polypeptides, whereas the other, which is missing in the RhD-negative condition, encodes the D protein that carries the major antigen of the RH system. Recently, different splicing isoforms transcribed from the CcEe gene were isolated. We report now the characterization of two other Rh clones, RhII and RhXIII, generated by alternative choices for poly(A) addition sites that were identified as the RhD gene transcripts. That these cDNAs represented the RhD messenger and that the previously described Rh clones were derived from the CcEe gene was demonstrated by amplification of RhII/XIII sequences only from D-positive genomes and by cloning and sequencing of D- and CcEe-specific gene fragments. The predicted translation product of the RhD mRNA is a 417-amino acid protein (M(r) = 45,500) that exhibited a similar membrane organization with 13 bilayer-spanning domains compared with the polypeptide encoded by the CcEe gene. The D and Cc/Ee polypeptides differ by 36 amino acid substitutions (8.4% divergence), but the NH2- and COOH-terminal regions of the two proteins are well conserved. Similarly, five of the six cysteine residues of the Cc/Ee proteins were conserved in the D protein, including the unique exofacial cysteine, which is critical for antigenic reactivity. The sequence homology between the Cc/Ee and D proteins supports the concept that the genes encoding these polypeptides have evolved by duplication of a common ancestor gene.
authors.affiliation: Unité Institut National de la Santé, Institut National de Transfusion Sanguine, Paris, France.
authors.forename: C
authors.initials: C
authors.lastname: Le van Kim
authors.forename: I
authors.initials: I
authors.lastname: Mouro
authors.forename: B
authors.initials: B
authors.lastname: Chérif-Zahar
authors.forename: V
authors.initials: V
authors.lastname: Raynal
authors.forename: C
authors.initials: C
authors.lastname: Cherrier
authors.forename: J P
authors.initials: JP
authors.lastname: Cartron
authors.forename: Y
authors.initials: Y
authors.lastname: Colin
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D001853:Bone Marrow / Q000502:physiology; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015342:DNA Probes; D005796:Genes; D015698:Genomic Library; D006801:Humans; D046911:Macromolecular Substances; D008961:Models, Structural; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D012333:RNA, Messenger / Q000235:genetics; D012204:Rh-Hr Blood-Group System / Q000235:genetics*
chemical_list: D015342:DNA Probes; D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D012204:Rh-Hr Blood-Group System; D004247:DNA