title: Human D(IIIa) erythrocytes: RhD protein is associated with multiple dispersed amino acid variations.
abstract: As a partial D antigen of the Rh blood group system, the D category IIIa phenotype occurs mainly in Blacks, but its molecular basis has not been defined. Here we describe studies of the D category D(IIIa) and VS+ red blood cells (RBC) from two unrelated probands by Southern blot, cDNA PCR, and nucleotide sequencing. Rh haplotyping by Sph I restriction fragment length polymorphisms indicated that the two probands carried Dce/dCe and Dce/DcE genotypes, respectively. Sequence analysis of Rh cDNAs showed that their erythroid cells expressed both D and CE transcripts. Nevertheless, the D transcripts were found to contain four nucleotide changes scattered in three exons: nt455 A-to-C (exon 3), nt602 C-to-G (exon 4), nt 654 C-to-G (exon 5), and nt667 T-to-G (exon 5). These variations resulted in the following amino acid substitutions characteristic of RhCE polypeptides: 152 Asn-to-Thr, 201 Thr-to-Arg, 218 Ile-to-Met, and 223 Phe-to-Val. The 152Thr and 223Val residues were predicted to reside in proximity to the third and fourth extracellular loops, respectively. Together, these results establish a correlation of the four amino acid changes in the RhD protein with the expression of D(IIIa) as a partial D antigen on the RBC membrane. Since the varied nucleotides identified in D(IIIa) all pre-exist in CE, they are likely to have originated from CE by templated micro-conversion event(s). The identification of a specific nt736 C-to-G transversion in CE in the two probands suggests that 245Val may involve the expression of VS antigen.
authors.affiliation: Laboratory of Biochemistry and Molecular Genetics, Lindsley F. Kimball Research Institute, New York Blood Center, NY 10021, USA.
authors.forename: C H
authors.initials: CH
authors.lastname: Huang
authors.forename: Y
authors.initials: Y
authors.lastname: Chen
authors.forename: M
authors.initials: M
authors.lastname: Reid
mesh_terms: D000596:Amino Acids / Q000032:analysis; D044383:Black People / Q000235:genetics; D015139:Blotting, Southern; D004247:DNA / Q000737:chemistry; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004912:Erythrocytes / Q000737:chemistry*; D005091:Exons; D005838:Genotype; D006238:Haploidy; D006801:Humans; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012204:Rh-Hr Blood-Group System / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D000596:Amino Acids; D012204:Rh-Hr Blood-Group System; C026410:Rho(D) antigen; D004247:DNA; D015247:Deoxyribonuclease HindIII; C046184:endodeoxyribonuclease PaeI; D015252:Deoxyribonucleases, Type II Site-Specific