RESUMEN
Description of three novel RAET1E/ULBP4 allele and promoter polymorphisms identified by sequence-based typing.
Asunto(s)
Alelos , Proteínas Portadoras/genética , Mutación del Sistema de Lectura , Antígenos de Histocompatibilidad Clase I/genética , Células Asesinas Naturales/inmunología , Proteínas de la Membrana/genética , Polimorfismo Genético , Regiones Promotoras Genéticas , Proteínas Portadoras/inmunología , Clonación Molecular , Codón sin Sentido , Exones , Femenino , Sangre Fetal/citología , Sangre Fetal/inmunología , Expresión Génica , Sitios Genéticos , Antígenos de Histocompatibilidad Clase I/inmunología , Humanos , Células Asesinas Naturales/citología , Masculino , Proteínas de la Membrana/inmunología , Subfamilia K de Receptores Similares a Lectina de Células NK/genética , Subfamilia K de Receptores Similares a Lectina de Células NK/inmunología , Análisis de Secuencia de ADNRESUMEN
We have now found a total of 15 individual MICB promoter sequences, varying by combination of 18 polymorphic positions within the MICB minimal promoter sequence. Sequence-based typing and cloning characterized the three new 5' promoter sequences as MICB-P13, MICB-P14 and MICB-P15.
Asunto(s)
Variación Genética , Antígenos de Histocompatibilidad Clase I/genética , Regiones Promotoras Genéticas , Alelos , Haplotipos/genética , Haplotipos/inmunología , Antígenos de Histocompatibilidad Clase I/inmunología , HumanosRESUMEN
In this study, we have characterized two novel polymorphism of the 5' promoter sequence of MICA gene, MICA-P13 and MICA-P14, by sequence-based typing and cloning.
Asunto(s)
Antígenos de Histocompatibilidad Clase I/genética , Regiones Promotoras Genéticas , Alelos , Haplotipos , Humanos , Datos de Secuencia Molecular , Polimorfismo de Nucleótido Simple , Análisis de Secuencia de ADNRESUMEN
The major histocompatibility complex (MHC) class I-related chain A (MICA) and B (MICB) are ligands for the natural killer group 2, member D (NKG2D) activating receptor expressed on natural killer (NK) cells, natural killer T (NKT) cells, CD8+ T cells and γδ T cells. Natural killer group 2, member D (NKG2D) ligand expression is stress-related and upregulated by infected or oncogenic cells leading to cytolysis. MICA and MICB genes display considerable polymorphism among individuals and studies have investigated allelic association with disease and relevance of MICA in transplantation, with variable success. It is now known that promoters of MICA and MICB are polymorphic with some polymorphisms associating with reduced expression. We sequenced International Histocompatibility Workshop (IHW) cell line DNA to determine promoter types and alleles encoded by exons 2-6. We found 8 of 12 known MICA promoter polymorphisms and although promoter P7 dominated, other promoters associated with the same allele. For example, MICA*002:01 had promoters P3, P4 or P7 and the common MICA*008:01/04 type had P1, P6 or P7. Similarly, we sequenced 8 of 12 known MICB promoter haplotypes. Some coding region defined MICB alleles had a single promoter, for example, MICB*002:01 and promoter P9, whereas the promiscuous MICB*005 allele had promoters P1, P2, P5, P6, P10 or P12. The results indicate potential for variation in expression of MICA and MICB ligands between individuals with the same allelic types. If differential expression by polymorphic MICA and MICB promoters is confirmed by functional studies, involvement of these genes in disease susceptibility or adverse transplantation outcomes may require knowledge of both promoter and allelic types to make meaningful conclusions.
Asunto(s)
Antígenos de Histocompatibilidad Clase I/genética , Células Asesinas Naturales/fisiología , Subfamilia K de Receptores Similares a Lectina de Células NK/agonistas , Regiones Promotoras Genéticas/genética , Citotoxicidad Inmunológica , Análisis Mutacional de ADN , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Haplotipos , Antígenos de Histocompatibilidad Clase I/metabolismo , Humanos , Ligandos , Polimorfismo Genético , Reino UnidoRESUMEN
Description of a novel RAET1E/ULBP4 allele characterized by sequence-based typing and cloning: RAET1E*011.
Asunto(s)
Alelos , Proteínas Portadoras/genética , Antígenos de Histocompatibilidad Clase I/genética , Proteínas de la Membrana/genética , Polimorfismo Genético , Secuencia de Bases , Clonación Molecular , Femenino , Humanos , Masculino , Datos de Secuencia MolecularRESUMEN
Discovery of three novel alleles of RAET1E/ULBP4 by sequence-based typing: RAET1E*008, RAET1E*009 and RAET1E*010.
Asunto(s)
Alelos , Proteínas Portadoras/genética , Etnicidad , Antígenos de Histocompatibilidad Clase I/genética , Proteínas de la Membrana/genética , Polimorfismo de Nucleótido Simple , Sustitución de Aminoácidos , Secuencia de Bases , Proteínas Portadoras/inmunología , Línea Celular , Cromosomas Humanos Par 6 , Exones , Sitios Genéticos , Antígenos de Histocompatibilidad Clase I/inmunología , Humanos , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Ligandos , Proteínas de la Membrana/inmunología , Datos de Secuencia Molecular , Tipificación Molecular , Subfamilia K de Receptores Similares a Lectina de Células NK/genética , Subfamilia K de Receptores Similares a Lectina de Células NK/inmunología , Isoformas de Proteínas/genética , Isoformas de Proteínas/inmunología , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido NucleicoRESUMEN
We report the identification of two novel major histocompatibility complex (MHC) class I-related chain A (MICA) alleles. MICA*054 has a nucleotide substitution of A to G at position 871 (codon 268), encoding an amino acid change of serine to glycine in the alpha-3 domain. MICA*056 has a nucleotide substitution at position 758 of G to C resulting in the substitution of tryptophan for serine at codon 230, also in the alpha-3 domain.
Asunto(s)
Alelos , Sustitución de Aminoácidos/genética , Antígenos de Histocompatibilidad Clase I/genética , Secuencia de Bases , Exones/genética , Humanos , Datos de Secuencia Molecular , Alineación de SecuenciaRESUMEN
We identified an A*0114 allele in an Irish patient with an apparent A*0236, A*3601 type.
Asunto(s)
Antígenos HLA/genética , Antígenos HLA-A/genética , Alelos , Secuencia de Bases , Cartilla de ADN/química , Antígeno HLA-A1 , Antígeno HLA-A2 , Humanos , Linfocitos/inmunología , Datos de Secuencia Molecular , Polimorfismo Genético , Población BlancaRESUMEN
A new HLA-A*31 null allele results from addition of an extra C near the beginning of exon 4 after a string of seven Cs.
Asunto(s)
Alelos , Antígenos HLA-A/genética , Secuencia de Bases , Exones , Humanos , Datos de Secuencia Molecular , Alineación de SecuenciaRESUMEN
We report here the full-length sequence of a novel HLA-A*0301 allele, A*03010103, which differs from A*03010101 by a single nucleotide substitution (G>T) at position 492 within intron 2. The variant was originally identified by Reference Strand-mediated Conformational Analysis (RSCA) and was confirmed by cloning and sequencing. The difference in RSCA mobility between A*03010101 and A*03010103 demonstrates the sensitivity of RSCA to detect single nucleotide polymorphisms.
Asunto(s)
Antígenos HLA-A/genética , Intrones , Secuencia de Bases , Antígenos HLA-A/inmunología , Antígeno HLA-A3 , Humanos , Datos de Secuencia MolecularRESUMEN
A novel human leucocyte antigen-A*02 (HLA-A*02) allele was detected by reference strand-mediated conformation analysis (RSCA) of a DNA sample from a Tarahumara individual. Direct sequencing of HLA-A locus polymerase chain reaction products identified a mutation in one of the alleles. Cloning and sequencing confirmed the presence of a new allele, A*0257 which differed from A*0206 by two nucleotides at positions 355 and 362, inducing changes in residues 95 and 97, respectively, within the peptide-binding site. Those changes suggest that allele A*0257 may have resulted from an intralocus recombination event.
Asunto(s)
Alelos , Antígenos HLA-A/genética , Mutación , Secuencia de Bases , Antígenos HLA-A/inmunología , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADNRESUMEN
Currently most available HLA-A, -B and -C DNA sequences cover exons 2 and 3 with a limited number extending to include other exons and introns. We have developed a method for the accurate determination of full-length genomic DNA sequences for HLA-A, -B and -C alleles. The method involves cloning of PCR amplified full-length HLA genes to separate alleles at heterozygous loci. The approach avoids any ambiguities from sequencing heterozygous PCR products directly and also avoids ambiguities from sequencing overlapping PCR products to achieve full-length sequence. To date we have sequenced full-length genomic sequences from representatives of all the major HLA-B and -C allele groups.
Asunto(s)
Antígenos HLA-B/genética , Antígenos HLA-C/genética , Alelos , Secuencia de Bases , Clonación Molecular , Cartilla de ADN , Antígenos HLA-B/análisis , Antígenos HLA-C/análisis , Humanos , Internet , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
We report the definition of an HLA class I null allele that has been identified within the B35 group by a combination of serological and molecular typing. This allele, which has been named B*3540N, was detected in a French, potential unrelated hematopoietic stem cell donor of unknown ethnic origin, selected as a probable match for an Irish patient. The presence of the null allele was initially determined by the absence of B35 reactivity by serological typing, in contrast to positive reactions by PCR-SSP and PCR-SSO typing. Subsequent sequencing of clones containing the full genomic sequence of the B*35 allele identified a single nucleotide deletion within exon 4 which resulted in the introduction of a stop codon downstream within exon 4.
Asunto(s)
Antígenos HLA-B/genética , Indenos/farmacología , Piridinas/farmacología , Alelos , Secuencia de Bases , Humanos , Indenos/química , Datos de Secuencia Molecular , Piridinas/químicaRESUMEN
Since the development of the polymerase chain reaction, most HLA class II allele sequencing has been exclusively focused on the highly polymorphic exon 2. We present here the full cDNA sequences of two HLA-DRB1 alleles, DRB1*0402 and DRB1*11041, both of which were previously only available as partial sequences. HLA-DRB1*11041 was found to be completely homologous to DRB1*11011 in exons 1, 3, 4, 5 and 6 and HLA-DRB1*0402 was found to be identical to DRB1*04011 in exons 1, 3, 4, 5 and 6.
Asunto(s)
Antígenos HLA-DR/genética , Alelos , Secuencia de Bases , ADN Complementario/genética , Exones , Cadenas HLA-DRB1 , Humanos , Datos de Secuencia MolecularRESUMEN
A novel polymorphism was identified in a B*15 allele. B*1566 possesses a nucleotide substitution of C to G at nucleotide 272. This polymorphism encodes an amino acid difference from serine in B*1501101 to cysteine in B*1566 at residue 67. Residue 67 is a constituent of the B pocket and is situated on the alpha1 helix facing into the groove. This mutation may have arisen through interallelic recombination as it has been seen in other B*15 alleles and is also present in most B*14, B*27, B*38, B*39 alleles and in B*7301.
Asunto(s)
Antígenos HLA-B/genética , Polimorfismo Genético , Sustitución de Aminoácidos/genética , HumanosRESUMEN
Two new HLA-C alleles have been identified by reverse line blot assay and direct sequencing of polymerase chain reaction (PCR) products. The first polymorphism encodes an amino acid change from lysine in Cw*12022 to asparagine in Cw*1208 at residue 66. The second polymorphism encodes two amino acid changes from isoleucine in Cw*03031 to threonine in Cw*0313 at residue 93 and isoleucine to leucine at residue 94. The functional significance of these polymorphisms on peptide-binding and/or T-cell recognition is unknown.
Asunto(s)
Alelos , Antígenos HLA-C/genética , Genes MHC Clase I , Prueba de Histocompatibilidad , Humanos , Modelos Genéticos , Datos de Secuencia Molecular , Síndromes Mielodisplásicos/genética , Síndromes Mielodisplásicos/patología , Análisis de Secuencia de ADN , Análisis de Secuencia de ProteínaRESUMEN
Sequencing studies of HLA class II molecules have been focused almost exclusively on the highly polymorphic exon 2. In this study the complete cDNA sequence of four alleles of the DR14 lineage (DR52 group) are reported for the first time. The HLA-DRB1*1402 and *1403 sequences were shown to be identical to the previously determined DRB1*13011 sequence, also of the DR52 group, in exons 1, 3, 4, 5 and 6. HLA-DRB1*14011 and *1404 were identical to DRB1*13011 in exons 1, 4, 5 and 6 sequences while they showed specific features within their exon 3 sequence. Both alleles showed a synonymous substitution at the third base of codon 114. However, DRB1*14011 also has a non-synonymous substitution at the first base of codon 112 which results in a histidine to tyrosine substitution. This is a novel substitution as Histidine 112 is conserved in all known HLA class II B genes.
Asunto(s)
Alelos , Antígenos HLA-DR/genética , Secuencia de Bases , Células Cultivadas , ADN Complementario/análisis , Exones , Cadenas HLA-DRB1 , Humanos , Datos de Secuencia Molecular , Polimorfismo Genético , Análisis de Secuencia de ADNRESUMEN
The MHC class I chain-related (MIC) gene family constitutes an interesting genetic group that is related to major histocompatibility complex (MHC) class I genes and is located within the MHC. The MIC gene products, MICA and MICB, have similar structures to HLA class I molecules. So far over 50 MICA alleles have been reported, which suggests that this genetic system is highly polymorphic. In order to investigate further the extent of MICA polymorphism we have studied exons 2-5 of the MICA gene in over 200 homozygous and heterozygous cell lines. Altogether we have identified 11 new MICA alleles and report 13 new nucleotide variations, one in exon 2, four in exon 3, four in exon 4, two in intron 1, one in intron 4 and one (a deletion) in exon 4. Eight of the 10 exonic variations are non-synonymous. The deletion in exon 4 leads to a frame-shift mutation and the introduction of a repeat of 12 leucine residues encoded by the microsatellite in exon 5. This study provides further evidence that the MICA gene is highly polymorphic. In contrast to MHC class I molecules, the polymorphic sites in MICA are predominantly within the alpha2 and alpha3 domains. The distribution of synonymous and non-synonymous substitutions suggests that there is selection for the polymorphic positions, which therefore define potential functional sites in the protein. We were also able to determine the association between MICA and HLA-B alleles in a number of homozygous cell lines bearing extended haplotypes.
Asunto(s)
Polimorfismo Genético , Alelos , Secuencia de Bases , ADN , Exones , Humanos , Repeticiones de Microsatélite , Datos de Secuencia MolecularRESUMEN
A novel polymorphism was identified in a B*07 allele. B*0722 possesses a unique nucleotide substitution at nucleotide 481. This polymorphism encodes an amino acid difference from aspartic acid in B*07021 to asparagine in B*0722. The functional significance of this polymorphism on peptide binding and/or T-cell recognition is unknown.