Molecular Basis of the Kell-null Phenotype

A MUTATION AT THE SPLICE SITE OF HUMAN KEL GENE ABOLISHES THE EXPRESSION OF KELL BLOOD GROUP ANTIGENS*

Lung-Chih YuDagger , Yuh-Ching TwuDagger , Ching-Yi ChangDagger , and Marie LinDagger §

From the Dagger  Transfusion Medicine Laboratory, Department of Medical Research, and the § Immunohematology Reference Laboratory, Mackay Memorial Hospital, Taipei 251, Taiwan

Received for publication, October 30, 2000, and in revised form, December 12, 2000


    ABSTRACT
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ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES

The Kell blood group system is polymorphic, and 23 antigens have been defined to date. The Kell antigens are located on a single red cell transmembrane glycoprotein, encoded by the 19 exons of the KEL gene. The different Kell phenotypes result from point mutations leading to amino acid changes in the Kell glycoprotein. An unusual phenotype, which is defined as the complete lack of all of the Kell antigens, has been identified and designated as the Kell-null or Ko phenotype. The coding region of the KEL gene of the Ko individual showed a normal KEL2/KEL4/KEL7 gene sequence; nevertheless, a G to C mutation at the splice donor site (5' splice site) of intron 3 was found to be present as a homozygote in the individual. The mutation destroys the conserved GT sequence of the splice donor site. Reverse transcription-polymerase chain reaction analysis showed the absence of the complete KEL mRNA. Instead, a major transcript with the exon 3 region skipped was found. The exon 3 of the KEL gene encodes the transmembrane domain of the Kell glycoprotein, and a transcript without exon 3 is predicted to have a premature stop codon that abolishes the translation of C-terminal segment. The segment contains all of the known positions responsible for characterizing different Kell antigens, and this explains the lack of all Kell antigens in Ko red cells.


    INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES

Beginning with the first discoveries of the K antigen by Coombs et al. in 1946 (1) and of its antithetical k antigen by Levine et al. in 1949 (2), the human Kell blood group system has been shown to be highly polymorphic. A series of the Kell antigens express antithetically with high and low frequencies, including KEL2 (k)/KEL1 (K), KEL4 (Kpb)/KEL3 (Kpa)/KEL21 (Kpc), KEL7 (Jsb)/KEL6 (Jsa), KEL11/KEL17, and KEL14/KEL24. The others have no known antithetical partner and are called para-Kell antigens. To date, 23 Kell antigens have been defined (3-7). Some Kell antigens exhibit ethnic specificity, for example, the K and Jsa antigens are characteristics of Europeans and Blacks, respectively. The former is found among the populations of European countries and varies in frequency from 4 to 9%, but is virtually absent in Orientals. Jsa, a low frequency antigen in general, has a high incidence of about 20% in Blacks.

The Kell blood group antigens are potent immunogens aside from the A and B antigens of the ABO system and the D antigen of the Rh system. Anti-K antibody has been identified as causing severe hemolytic disease of the newborn and severe hemolytic transfusion reactions. The Kell system antibodies have been further shown to suppress erythropoiesis, and recently they were also demonstrated to suppress megakaryopoiesis (8). Thus, in the field of transfusion medicine, Kell system is considered to be most important after the ABO and Rh systems, especially in Western countries.

The Kell and para-Kell antigens are located on a single transmembrane glycoprotein with a molecular mass of 93 kDa (9). The Kell polypeptide is covalently linked to another blood group protein, Kx, by a disulfide bond (10, 11). A series of molecular studies, accomplished by Lee et al. (12) have revealed the primary structure of the Kell protein and genomic organization of the KEL gene (13) by cDNA and genomic DNA cloning, respectively, and consequently, the molecular bases for most polymorphic Kell phenotypes have been identified (Refs. 14-17; reviewed in Refs. 18-20). The KEL cDNA sequence predicts a 732-amino acid polypeptide with a single type II transmembrane domain (residues 48-67) and a cytoplasmic region made up of the N-terminal 47 amino acids. The extracellularly located C-terminal 665-amino acid segment is where all of the known positions responsible for characterizing different Kell phenotypes are found. The coding region of the KEL gene is spread over 19 exon regions spanning more than 20 kilobase of genomic DNA and is on chromosome 7q33. The entire transmembrane region is encoded by exon 3.

The Kell polypeptide has been shown to possess sequence and structural similarity to a family of mammalian neutral endopeptidases, and recently, it has been further shown to have proteolytic activity (21) and process the big endothelin-3 to yield endothelin-3, a potent bioactive peptide with multiple biological roles. However, the physiological role of Kell protein is still obscure, although the protein is distributed wider than was previously thought, not only in erythroid tissues but also in nonerythroid tissues, such as testis, skeletal muscle, etc. (22).

Expression of the Kell antigens can be depressed in certain instances, such as in the McLeod (23, 24), Gerbich-negative, Kpa, and Kmod phenotypes (4, 5). In 1957, Chown et al. (25) first described an unusual Kell-null (Ko) phenotype, which is defined as the complete lack of all known Kell antigens on the red cells. Although it is very rare, the Ko phenotype is widely distributed and has been identified in Europeans, Japanese, Africans, and Indians (3). The absence of the Kell antigens on these Ko cells was shown to be due to lack of the protein on the red cell membrane, because polyclonal antibodies against the Kell protein failed to detect any Kell protein on the Ko red cell membrane by Western blotting (23, 24). Although there have been molecular studies on this phenotype, the molecular basis for this mutation is still unknown.

In this paper we analyze the molecular basis of the Ko phenotype of a Chinese individual. A G to C substitution at the splice donor site (5' splice site) of intron 3 of the KEL gene was found to be present as a homozygote in the Ko individual and demonstrated to be responsible for the complete lack of all Kell antigens in the Ko red cells.

    EXPERIMENTAL PROCEDURES
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ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES

The Ko Case-- A Chinese individual with a group O phenotype and a negative direct antiglobulin test was noticed because he carried an alloantibody against a high frequency antigen during a pretransfusion test. The serum gave a positive reaction with all cells tested, including the panel cells of the DiaPanel (DiaMed, lot number 1621.70), 30 group O Taiwanese donor cells, Di(a+b-) cells and Vel negative cells, either by the manual Polybrene method or the LISS (low ionic strength saline) antiglobulin test. Blood group typing of the red cells of this individual was R1r (or R1R0), K-, k-, Kpa-, Kpb-, Jsa-, Jsb-, Fya+, Fyb-, Jka+, and Jkb+ (tested by FDA-approved commercial antisera except for anti-Jsa, which was a gift from Dr. J. Molds of Ortho Diagnostic Systems). The red cells showed a negative reaction with two anti-Ku sera obtained through the Serum Cells and Rare Fluids International Immunohematological Exchange Group. These two sera were from group A, Ko persons in the United States and Germany.

Sequence Analysis of the KEL Gene-- Genomic DNAs of the Ko and other normal individuals were prepared from peripheral white blood cells using the QIAamp Blood Kit (Qiagen).

The 19 exon regions of the KEL gene of the Ko individual were divided into nine segments and amplified by polymerase chain reaction (PCR).1 PCR primers used in this study were designed based on the papers of Lee et al. (12, 13) and are listed in Table I. 100 ng of genomic DNA and 15 pmol of each primer were combined in 25 µl of PCR buffer containing 0.2 mM of dNTP and 0.5 unit of Taq polymerase (Promega). The PCR program consisted of 5 min at 94 °C followed by 30 cycles of 1 min at 94 °C, 30 s at 50 °C, and 1 min at 72 °C. Sequences were directly determined from the PCR products using the BigDye Terminator Cycle Sequencing Kit (Applied Biosystems). The KEL gene sequence of the exon 3 to exon 4 region, amplified by the primer set, inF3 and inR4, of an individual with a normal KEL2 (KEL:-1,2 or K-k+) phenotype was also analyzed to inspect the exon 3-intron 3 junction and to allow comparison with that of the Ko individual.

                              
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Table I
Oligonucleotide primers used for PCR and RT-PCR amplification

PCR-Restriction Fragment Length Polymorphism (RFLP) Analysis-- The G to C nucleotide substitution identified at the splice donor site of intron 3 of the Ko gene creates a DdeI recognition sequence (CTNAG). PCR-based RFLP analysis was carried out to analyze this nucleotide position. The exon 3 to exon 4 region of the KEL gene was amplified by inF3 and inR4 primers as above, and the 624-bp PCR product was digested by the DdeI restriction endonuclease and then analyzed using a 3.0% agarose gel. In addition to the DNAs from Ko and the normal KEL2 individuals, another 66 genomic DNAs obtained from randomly selected individuals were also subjected to the PCR-RFLP analysis to measure the incidence of this G to C mutation in the KEL genes of the general population.

Reverse Transcription (RT)-PCR Analysis-- Total RNA were isolated from peripheral blood cells using the QIAamp Mini RNA Blood Kit (Qiagen). The One-Step RT-PCR Kit (Qiagen) was employed. 0.4 µg of total RNA and 15 pmol of each primer, exFg and exRa (Table I), which anneal to sequences within exon 1 and exon 19 regions, respectively, were combined in a total volume of 25 µl containing 0.4 mM of dNTP and the RT-PCR enzyme mix, following the manufacturer's instructions. The RT-PCR program consisted of 30 min at 50 °C for RT and then 15 min at 95 °C followed by 30 cycles of 1 min at 94 °C, 30 s at 60 °C, and 2 min at 72 °C. Nested PCR using primer sets of exFd and exRe or exFb and exRf (Table I) was then performed to sample from exon 2 to exon 8 or from exon 1 to exon 19 cDNA regions, respectively, using 1 µl of the RT-PCR product as a template. The program for the nested PCRs consisted of 5 min at 94 °C and then 30 cycles of 1 min at 94 °C and 2.5 min at 72 °C. The exon 2 to exon 8 product was analyzed by 2.5% agarose gel electrophoresis and sequencing. The nested amplified exon 1 to exon 19 products from three normal individuals were analyzed to reveal the general KEL transcript structure.

    RESULTS
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ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES

Identification of a Point Mutation at the Splice Donor Site of Intron 3 of the Ko Gene-- The 19 exon regions of the KEL gene were divided into 9 segments and amplified from genomic DNA by PCR, as described above. The amplified products were directly sequenced to analyze the coding regions. The entire coding sequence of the KEL gene of the Ko individual was shown to be identical with that of a normal KEL2/KEL4/KEL7 gene. No substitution, deletion, or insertion was identified. However, a nucleotide substitution, G to C, was found at the splice donor site of the intron 3 (Fig. 1, right panel). The mutation destroys the consensus sequence (GT) for the splice donor site, and the sequencing result suggests a homozygous state for the mutation in the Ko individual. Other splice donor and acceptor sites of the KEL gene of the Ko individual were demonstrated to have the conserved GT and AG sequences, respectively. A parallel sequencing analysis on the exon 3-intron 3 junction of the KEL gene of an individual with a normal KEL2 phenotype demonstrated the consensus GT sequence (Fig. 1, left panel).


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Fig. 1.   Sequencing results of the exon 3-intron 3 junction of KEL genes of Ko and normal KEL2 individuals. Genomic DNAs were purified from an individual with a normal KEL2 (KEL:-1,2 or K-k+) phenotype and the Ko individual. The exon 3 to exon 4 regions of the KEL genes were amplified by PCR. The products were directly sequenced using the PCR primer inF3 as a sequencing primer. The sequencing results of the exon 3-intron 3 regions of the normal KEL2 (left panel) and the Ko (right panel) samples are shown. The G to C change at the splice donor site is underlined. A dashed line indicates the new DdeI recognition sequence (CTNAG) created by the G to C change.

Because the G to C substitution created a DdeI restriction site (Fig. 1, right panel), a PCR-RFLP analysis was used to study this nucleotide position. DdeI digestion of the 624-bp PCR product, encompassing the exon 3 to exon 4 region of the KEL gene amplified from the normal KEL2 individual, yielded 330-, 219-, and 75-bp fragments (Fig. 2, A and B, lane 1). The 330-bp fragment generated from the Ko individual was further cleaved into 170- and 160-bp fragments (Fig. 2, A and B, lane Ko). The pattern in the lane Ko also suggested that the Ko individual is homozygous for the G to C mutation and agrees with the result from the direct sequencing. However, another possibility that he has a haploid mutant genotype at the KEL locus cannot be excluded.


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Fig. 2.   PCR-RFLP analysis of the splice donor site change in KEL gene. A, schematic of DdeI cleavage sites of the PCR-RFLP system. The 624-bp PCR product of the exon 3 to exon 4 region amplified from a normal KEL2 gene yields 219-, 75-, and 330-bp fragments, after digested with DdeI restriction endonuclease. The G to C nucleotide substitution at the splice donor site of intron 3 of the Ko gene creates an additional DdeI site (asterisk), which further divides the 330-bp fragment into 170- and 160-bp fragments. B, PCR-RFLP analysis results obtained from the normal KEL2 and the Ko individual. The KEL2 (lane 1) and the Ko (lane Ko) individuals were subjected to the analysis. The DdeI-cleaved products were analyzed by 3.0% agarose gel electrophoresis. Molecular mass standards (100-bp ladder) are in lane M. The result suggests that the Ko individual is most likely homozygous for the G to C mutation and that the substitution is absent in the normal KEL2 individual.

Genomic DNAs from another 66 randomly selected individuals were subjected to the PCR-RFLP analysis. The results (data not shown) demonstrated that no other KEL allele possessed this nucleotide substitution in the individuals tested, suggesting that the mutation is rare in the general population.

Absence of Correct KEL mRNA Transcript in the Ko Individual-- To reveal the effect on the KEL transcript structure exerted by the mutated splice donor site in the Ko gene, the KEL mRNA of the Ko individual was analyzed by RT-PCR and compared with mRNA obtained from 11 randomly selected common individuals.

The first RT-PCR sampled the exon 1 to exon 19 region of the KEL cDNA, and the following nested PCR amplified the exon 2 to exon 8 region. All of the 11 normal individuals generated a product with expected size of 909 bp (six of them are shown in lanes 1-6 of Fig. 3), and direct sequencing demonstrated the complete exon 2 to exon 8 structure. The 909-bp fragment was shown to be the major product in all of the 11 samples, except for one individual shown in lane 2, where it was minor. Nevertheless, the 909-bp fragment was absent when the RNA of the Ko individual was amplified, but a major product with a smaller size of 767 bp was obtained instead (Fig. 3, lane Ko). Sequencing analysis showed the DNA fragment consisting of sequence from exon 2 to exon 8 but without the 142-bp sequence of the exon 3 region.


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Fig. 3.   KEL transcript structures of the Ko and common individuals analyzed by RT-PCR. Total RNAs were purified from peripheral blood cells of the Ko and randomly selected individuals. One step RT-PCR was performed using the primers annealing to the sequences of exon 1 and exon 19 regions, and then nested PCR was employed to sample the exon 2 to exon 8 region of KEL cDNA. The nested PCR products were analyzed by 2.5% agarose gel electrophoresis. Molecular mass standards (100-bp) ladder are in lane M. The upper major band (909 bp, indicated by an asterisk) was demonstrated to have a complete exon 2 to exon 8 sequence by sequencing analysis and was present in all of the 11 normal individuals. Results obtained from six normal individuals are shown in lanes 1-6. However, the complete product was absent in the total RNA purified from the Ko individual (lane Ko). The lower major band (767 bp, indicated by two asterisks), consisting of the sequence of exon 2 to exon 8 but without exon 3 region, was the major product from the Ko individual. In the Ko sample, a minor transcript of exon 2 to exon 8 region with exons 3 and 6 removed was also produced (the 620-bp band). Transcripts without the exon 3 region, in addition to the complete form, were also present in some of the normal individuals (lanes 2 and 6) and are believed to result from alternative splicing (see "Discussion"). The minor band at approximately 550 bp in lane 6 is believed to amplified from a transcript lacking exons 3, 6, 7, and 9. Other minor products from the Ko sample and the minor band of about 350 bp in lane 4 are artifacts.

The result proved the complete lack of the correct KEL mRNA transcript and the presence of a major transcript with the exon 3 region skipped in the Ko individual. Obviously the transcript without exon 3 results from the mutation of G to C present in the Ko gene, which destroys the splice donor consensus of intron 3. The 620-bp minor product obtained from the Ko RNA was shown to have a structure lacking exon 3 and exon 6; other minor bands were found to be artifacts (Fig. 3, lane Ko).

In addition to the complete KEL transcript, a 767-bp product devoid of the exon 3 region was also found in four of the 11 normal individuals (two of the four are in lanes 2 and 6 of Fig. 3). Further studies showed that several forms of the KEL transcripts, in addition to the complete structure, could be demonstrated in the peripheral blood cell RNA of these normal individuals. These different transcripts are believed to result from alternative splicing.

    DISCUSSION
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ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES

The Kell epitopes are located on the single Kell polypeptide made up of 732 amino acid residues. The amino acid positions responsible for 18 of the known 23 antigens, among which 11 belong to five antithetical sets, have been revealed, and have been shown to localize extracellularly. All of the molecular bases have been shown to result from point mutations leading to amino acid substitution in the KEL gene. The positions of the remaining five epitopes have not yet been identified but are believed to lie in the Kell glycoprotein too.

The Kell-null or Ko phenotype, which represents the complete lack of the expression of all Kell antigens, although known for more than 40 years, has not had its molecular basis elucidated up to the present. The entire coding sequence of the KEL gene of our Ko case demonstrated a normal KEL2/KEL4/KEL7 gene sequence, but a nucleotide substitution of G to C identified at the splice donor site of intron 3 was found in the Ko individual. The results of the sequencing and PCR-RFLP analyses suggested that he is most likely homozygous for the mutation; however, the possibility that he has a haploid mutant genotype at the KEL locus should not be excluded without further analysis. The substitution destroys the conserved sequence of a splice donor site. RT-PCR analysis further proved that exon 3 was absolutely absent in the transcripts from the Ko gene. Thus, the destroyed donor splicing consensus makes the exon 3 region become a pseudoexon and results in the removal of exon 3 during KEL mRNA processing (Fig. 4).


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Fig. 4.   Diagrammatic representation of the molecular basis for the Ko allele. The mutated splice donor site (from GT to CT) of intron 3 of the Ko allele makes the exon 3 become a pseudoexon and causes the removal of the exon during KEL mRNA processing.

Three different consequences of aberrant mRNA splicing because of splice junction mutations have been observed: exon skipping, utilization of cryptic splice site, and intron retention. Data bases of aberrant splicing mutations in human (26) and mammalian (27) genes show that exon skipping is most frequently observed in aberrant splicing resulting from splice site mutation, whereas intron retention seldom occurs. The "exon definition model" proposed by Robberson et al. (28) gives a possible explanation for the skipping of exon adjacent to mutated splice site. Following the model to describe the normal mRNA maturation, the initial step of splicesome formation is exon definition, in which factors, including a variety of small nuclear ribonucleoproteins and associated proteins, binding to the splice acceptor site (3' splice site) followed by a search for and then the binding of factors to a downstream splice donor site. Factors at splice sites communicate across exons to define exon regions. After exon recognition, the splicing machinery cleaves introns and religates the coding sequences. In the absence of the consensus splice donor site, the splicing machinery searches for a neighboring best fit site, a cryptic site. If no acceptable site can be found in the vicinity of the authentic splice site, the splicesome complex becomes unstable resulting in insufficient preservation of exon region, and exon skipping seems to occur (28). The presence or absence of suitable cryptic splice site in the vicinity is an important factor in determining which event, skipping or cryptic site usage, occurs (26). As for the Ko gene, the splice donor site of intron 3 is destroyed, and obviously possible neighboring cryptic splice sites are not recognized. Thus, the whole exon 3 is skipped. Many examples of aberrant splicing in human genes and thus leading to genetic diseases have been demonstrated (26, 29-37). Not only the mutations at the most conserved GT sequence of a splice donor site but also those at the nearby conserved region can cause aberrant splicing (26, 30, 32, 36, 37). Many of them result in exon skipping. However, some cases represent combinations of different events, exon skipping and cryptic splice site usage (29, 37), alone and together with intron retention (33).

A KEL mRNA with the exon 3 skipped results in a reading frameshift, and the predicted polypeptide has an altered amino acid sequence and is truncated at C-terminal end by a premature stop codon (Fig. 5). The transmembrane segment of the Kell glycoprotein (residues 48-67, underlined in Fig. 5), which anchors the protein appropriately to the cell membrane, is encoded by exon 3, and of the 732-amino acid residues of Kell polypeptide, only the cytoplasmic N-terminal 27 amino acids are expressed correctly from the Ko gene (Fig. 5, right panel). The C-terminal segment is lost, and this is where all of the known Kell epitope positions are found. This explains the loss of all the Kell antigens in the Ko individual. Previous investigations using Western blotting analysis indicated the absence of Kell protein on the Ko red cells (23, 24), giving rise to the absence of all Kell epitopes. The molecular basis of the Ko phenotype presented here meets the observation.


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Fig. 5.   Comparison of the amino acid sequences predicted from the complete KEL mRNA and from the Ko transcript with exon 3 skipped. A normal KEL mRNA predicts a polypeptide of 732 amino acids (left panel) with a cytoplasmic region of N-terminal 47 amino acids, a transmembrane segment of residues 48-67 (underlined) encoded by exon 3, and a bulk of extracellularly located C-terminal 665 amino acids. The amino acid sequence predicted from the major Ko transcript (right panel) lacks the transmembrane domain and loses all of the extracellularly located C-terminal region where reside all of the known positions responsible for the different Kell antigens. The amino acid sequence of the Ko polypeptide as it differs from a normal Kell protein is in lowercase letters.

The correct KEL mRNA is absent, and instead, a transcript devoid of exon 3 region is the major form in the RNA of Ko cells. However, the form without exon 3 region was also present in four of the 11 normal individuals tested, although none of them had the G to C substitution in their KEL genes. Structures of the KEL transcripts of three of these normal individuals were further analyzed by sequencing the nested amplified RT-PCR products that encompassed the exon 1 to exon 19 cDNA region. In addition to the complete exon 1-19 transcript, KEL transcripts without exon 3, without exons 3 and 6, and without exons 3, 6, 7, and 9 were identified. The results suggest that there is frequent alternative splicing during the processing of the KEL mRNA, and exon 3 and exon 6 seem to be most commonly skipped.

Some mutations in the KEL gene can result in the weakening of Kell antigen expression. The Kpa phenotype has been known to be associated with a weakened expression of other Kell antigens. The missense mutation responsible for Kpa phenotype has been shown to cause aberrant transport of the Kell glycoprotein in Golgi compartment and consequently a reduction of the total amount of the protein at cell surface (38). A reduced quantity of Kell glycoprotein has also been demonstrated at the surface of Kmod cells, a rare phenotype with very weak Kell antigens. A different amino acid change was suggested to connect with the phenotype (39). Other missense mutations in the KEL gene produce the polymorphic antigens of the Kell blood group system. However, no nonsense mutation, deletion, or insertion in the KEL gene resulting in loss of part or all of the Kell antigens has been reported in the literature. This study shows that the abolition of the expression of all Kell antigens in the Ko phenotype is due to a base change at a splice site rather than the above changes.

However, it appears that different Ko cases might be caused by different molecular mechanisms. Lee et al. (19)2 have identified two unrelated Ko individuals who are heterozygous with one KEL allele possessing either a mutation at a consensus splice junction (which may be similar to our Ko case) or a nonsense mutation and with the other allele having point mutations leading to amino acid change.

    FOOTNOTES

* This work was partially supported by Grant NHRI-GT-EX89S601L from the National Health Research Institute and Grant NSC 89-2314-B-195-014 from the National Science Council, Taiwan.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Transfusion Medicine Lab., Dept. of Medical Research, Mackay Memorial Hospital, 45 Ming-San Rd., Tamshui, Taipei County 251, Taiwan. Tel.: 886-2-2809-4661, Ext. 2380; Fax: 886-2-2809-4679; E-mail: marilin@ms2.mmh. org.tw.

Published, JBC Papers in Press, December 27, 2000, DOI 10.1074/jbc.M009879200

2 S. Lee, D. Russo, and C. Redman, unpublished observation.

    ABBREVIATIONS

The abbreviations used are: PCR, polymerase chain reaction; RT, reverse transcription; RFLP, restriction fragment length polymorphism; bp, base pair(s).

    REFERENCES
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES

1. Cooms, R. R. A., Mourant, A. E., and Race, R. R. (1946) Lancet 1, 264-266
2. Levine, P., Backer, M., Wigod, M., and Ponder, R. (1949) Nature 109, 464-466
3. Redman, C. M., and Lee, S. (1995) in Blood Cell Biochemistry (Cartron, J.-P. , and Rouger, P., eds) , pp. 227-242, Plenum Press, New York
4. Daniels, G. (1995) Human Blood Groups , pp. 385-420, Blackwell Science Ltd., Oxford
5. Reid, M. E., and Lomas-Francis, C. (1997) The Blood Group Antigen FactsBook , pp. 175-209, Academic Press, San Diego, CA
6. Issitt, P. D., and Anstee, D. J. (1998) Applied Blood Group Serology , 4th Ed. , pp. 609-653, Montgomery Scientific Publications, Durham, NC
7. Schenkel-Brunner, H. (2000) Human Blood Groups: Chemical and Biochemical Basis of Antigen Specificity , 2nd Ed. , pp. 485-503, Springer-Verlag, Wien
8. Wagner, T., Bernaschek, G., and Geissler, K. (2000) N. Eng. J. Med. 343, 72[Free Full Text]
9. Redman, C. M., Avellino, G., Pfeffer, S. R., Mukherjee, T. K., Nichols, M., Rubinstein, P., and Marsh, W. L. (1986) J. Biol. Chem. 261, 9521-9525[Abstract/Free Full Text]
10. Russo, D., Redman, C., and Lee, S. (1998) J. Biol. Chem. 273, 13950-13956[Abstract/Free Full Text]
11. Russo, D., Lee, S., and Redman, C. (1999) Biochim. Biophys. Acta 1461, 10-18[Medline] [Order article via Infotrieve]
12. Lee, S., Zambas, E. D., Marsh, L., and Redman, C. M. (1991) Proc. Natl. Acad. Sci. U. S. A. 88, 6353-6357[Abstract]
13. Lee, S., Zambus, E., Green, E. D., and Redman, C. (1995) Blood 85, 1364-1370[Abstract/Free Full Text]; Correction (1998) 87, 4922
14. Lee, S., Wu, X., Reid, M., Zelinski, T., and Redman, C. (1995) Blood 85, 912-916[Abstract/Free Full Text]
15. Lee, S., Wu, X., Reid, M., and Redman, C. (1995) Transfusion 35, 822-825[CrossRef][Medline] [Order article via Infotrieve]
16. Lee, S., Wu, X., Son, S., Naime, D., Reid, M., Okubo, Y., Sistonen, P., and Redman, C. (1996) Transfusion 36, 490-494[Medline] [Order article via Infotrieve]
17. Lee, S., Naime, D. S., Reid, M. E., and Redman, C. M. (1997) Transfusion 37, 1117-1122[Medline] [Order article via Infotrieve]
18. Lee, S. (1997) Vox Sang. 73, 1-11[CrossRef][Medline] [Order article via Infotrieve]
19. Lee, S., Russo, D., and Redman, C. (2000) Transfus. Med. Rev. 14, 93-103[Medline] [Order article via Infotrieve]
20. Lee, S., Russo, D., and Redman, C. M. (2000) Semin. Hematol. 37, 113-121[Medline] [Order article via Infotrieve]
21. Lee, S., Lin, M., Mele, A., Cao, Y., Farmar, J., Russo, D., and Redman, C. (1999) Blood 94, 1440-1450[Abstract/Free Full Text]
22. Russo, D., Wu, X., Redman, C. M., and Lee, S. (2000) Blood 96, 340-346[Abstract/Free Full Text]
23. Redman, C. M., Marsh, W. L., Scarborough, A., Johnson, C. L., Rabin, B. I., and Overbeeke, M. (1988) Br. J. Haematol. 68, 131-136[Medline] [Order article via Infotrieve]
24. Jaber, A., Loirat, M.-J., Willem, C., Bloy, C., Cartron, J.-P., and Blanchard, D. (1991) Br. J. Haematol. 79, 311-315[Medline] [Order article via Infotrieve]
25. Chown, B., Lewis, M., and Kaita, K. (1957) Nature 180, 711[Medline] [Order article via Infotrieve]
26. Krawczak, M., Reiss, J., and Cooper, D. N. (1992) Hum. Genet. 90, 41-54[Medline] [Order article via Infotrieve]
27. Nakai, K., and Sakamoto, H. (1994) Gene 141, 171-177[CrossRef][Medline] [Order article via Infotrieve]
28. Robberson, B. L., Cote, G. J., and Berget, S. M. (1990) Mol. Cell. Biol. 10, 84-94[Medline] [Order article via Infotrieve]
29. Thierfelder, L., Watkins, H., MacRae, C., Lamas, R., McKenna, W., Vosberg, H.-P., Seldman, J. G., and Seidman, C. E. (1994) Cell 77, 701-712[Medline] [Order article via Infotrieve]
30. Winberg, J.-O., Hammami-Hauasli, N., Nilssen, Ø., Anton-Lamprecht, I., Naylor, S. L., Kerbacher, K., Zimmermann, M., Krajci, P., Gedde-Dahl, T., Jr., and Bruckner-Tuderman, L. (1997) Hum. Mol. Genet. 6, 1125-1135[Abstract/Free Full Text]
31. McVey, J. H., Boswell, E. J., Takamiya, O., Tamagnini, G., Valente, V., Fidalgo, T., Layton, M., and Tuddenham, E. G. D. (1998) Blood 92, 920-926[Abstract/Free Full Text]
32. Ohno, K., Berngman, J. M., Felice, K. J., Cornblath, D. R., and Engel, A. G. (1999) Am. J. Hum. Genet. 65, 635-644[CrossRef][Medline] [Order article via Infotrieve]
33. Schwarze, U., Starman, B. J., and Byers, P. H. (1999) Am. J. Hum. Genet. 65, 336-344[CrossRef][Medline] [Order article via Infotrieve]
34. Harteveld, C. L., Beijer, C., van Delft, P., Zanardini, R., Bernini, L. F., and Giordano, P. C. (2000) Br. J. Haematol. 110, 694-698[CrossRef][Medline] [Order article via Infotrieve]
35. Wyatt, J., Brennan, S. O., May, S., and George, P. M. (2000) Thromb. Haemostasis 84, 449-452[Medline] [Order article via Infotrieve]
36. Condino-Neto, A., and Newburger, P. E. (2000) Blood 95, 3548-3554[Abstract/Free Full Text]
37. Okubo, M., Horinishi, A., Suzuki, Y., Murase, T., and Hayasaka, K. (2000) Am. J. Med. Genet. 93, 211-214[CrossRef][Medline] [Order article via Infotrieve]
38. Yazdanbakhsh, K., Lee, S., Yu, Q., and Reid, M. E. (1999) Blood 94, 310-318[Abstract/Free Full Text]
39. Uchikawa, M., Onodera, T., Tsuneyama, H., Enomoto, T., Ishijima, A., Yuasa, S., Murata, S., Tadokoro, K., Nakajima, K., and Juji, T. (2000) Vox Sang. 78, O011 (abstr.)


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