GM02756

GM02756 LCL from B-Lymphocyte

Description:

SEVERE COMBINED IMMUNODEFICIENCY, AUTOSOMAL RECESSIVE, T CELL-NEGATIVE, B CELL-NEGATIVE, NK CELL-NEGATIVE, DUE TO ADENOSINE DEAMINASE DEFICIENCY
ADENOSINE DEAMINASE; ADA

Affected:

Yes

Sex:

Male

Age:

3 YR (At Sampling)

Overview

Repository

NIGMS Human Genetic Cell Repository

Subcollection

Heritable Diseases

Class

Disorders of Nucleotide and Nucleic Acid Metabolism

Biopsy Source

Peripheral vein

Cell Type

B-Lymphocyte

Tissue Type

Blood

Transformant

Epstein-Barr Virus

Sample Source

LCL from B-Lymphocyte

Race

Black/African American

Relation to Proband

proband

Confirmation

Molecular characterization after cell line submission to CCR

Species

Homo sapiens

Common Name

Human

Remarks

Clinically affected; deficient ADA activity in lymphoblasts and mononuclear cells; enzyme phenotypes: 6PGD=A, G6PD=B, Peptidases A,C,and D=1, Acid Aglucosidase=1, Neutral A-glucosidase C= 3-2; normal ADA mRNA level; donor subject is a compound heterozygote: one allele has a G>A transition at nucleotide 727 in exon 7 of the ADA gene [727G>A (alternately described as 632G>A)] resulting in a substitution of histidine for arginine at codon 211 [Arg211His (R211H)] and a second allele has a C>T transition at nucleotide 1081 in exon 11 of the ADA gene [1081C>T] resulting in a substitution of valine for alanine at codon 329 [Ala329Val (A329V)].

Characterizations

IDENTIFICATION OF SPECIES OF ORIGIN

Species of Origin Confirmed by Nucleoside Phosphorylase, Glucose-6-Phosphate Dehydrogenase, and Lactate Dehydrogenase Isoenzyme Electrophoresis

adenosine deaminase

Wiginton et al (J Biol Chem 257:3211-3217 1982) observed that this cell culture contained < 5% of normal ADA activity and immunoreactive protein. Adrian et al (Hum Genet 68:169-172 1984) reported that this ADA-deficient cell culture had normal relative levels of ADA mRNA. Sequence analysis of cDNA clones synthesized from ADA mRNA from this culture revealed a different point mutation in each allele that causes amino acid changes. In one allele amino acid 329 is changed from alanine to valine (GCG>GTG at position #1081). In the second allele amino acid 211 is changed from arginine to histidine (CGT>CAT at position 727). In addition to these point mutations each allele has a point mutation in the third base of a codon that does not alter the amino acid encoded (Ser-110 is TCT rather than TCC and Val-629 is GTA rather than GTG). Analysis of additional clones of each of these alleles showed unusual characteristics which suggest occasional aberrant splicing of ADA pre-mRNA (Akeson et al Proc Natl Acad Sci USA 84:59475951 1987). Akeson et al (J Biol Chem 263:16291-16296 1988) inserted the mutant ADA cDNA from this cell line into an expression vector and tested it in a transient expression system. ADA mRNA transcribed from the mutant vector failed to produce significant levels of enzymatically active ADA while ADA mRNA transcribed from a vector with the normal ADA cDNA did produce high levels of enzymatically active ADA. These authors also noted that the point mutation at amino acid no. 211 (Arg to His) resulting from a point mutation at base no. 727 (G to A) was also observed in another ADA-deficient cell line (GM02606) and the mutation in amino acid no. 329 (Ala to Val) resulting from a point mutation at base no. 1081 (C to T) had been observed for a different ADA-deficient cell line (GM02825). EC Number: 3.5.4.4; <5% activity.

GENE MAPPING & DOSAGE STUDIES - Y CHROMOSOME

PCR analysis of DNA from this cell culture gave a positive result with a primer for Yq11, DYS227.

Gene

ADA

Chromosomal Location

20q13.11

Allelic Variant 1

608958.0004; SEVERE COMBINED IMMUNODEFICIENCY, AUTOSOMAL RECESSIVE, T CELL-NEGATIVE, B CELL-NEGATIVE, NK CELL-NEGATIVE, DUE TO ADENOSINE DEAMINASE DEFICIENCY

Identified Mutation

ARG211HIS; Akeson et al. [J Biol Chem 263:16291 (1988)] found this change, a 632G-A transition in the ADA gene, resulting in the replacement of the arginine residue by histidine at codon 211, in cell line GM02606 and Akeson et al. [Proc Natl Acad Sci U S A 84: 5947 (1987)] found it in cell line GM02756.

Gene

ADA

Chromosomal Location

20q13.11

Allelic Variant 2

608958.0006; SEVERE COMBINED IMMUNODEFICIENCY, AUTOSOMAL RECESSIVE, T CELL-NEGATIVE, B CELL-NEGATIVE, NK CELL-NEGATIVE, DUE TO ADENOSINE DEAMINASE DEFICIENCY

Identified Mutation

ALA329VAL; In GM02756, Akeson et al. [Proc Natl Acad Sci U S A 84: 5947 (1987)] demonstrated 2 different mutant alleles: one was ala329 to val; the other was a 632G-A transition in the ADA gene, resulting in the replacement of the arginine residue by histidine at codon 211 (102700.0004). In addition, Akeson et al. [Proc Natl Acad Sci U S A 84: 5947 (1987)] and [J Biol Chem 263:16291 (1988)] found that cell line GM02825A was a genetic compound. One allele had an ala329-to-val change (102700.0006); the other allele had a point mutation from A to G in the 3-prime splice site of intron 3 (102700.0017), resulting in elimination of exon 4 from the mature mRNA.

Phenotypic Data

Remarks

Publications

Kouprina N, Pavlicek A, Noskov VN, Solomon G, Otstot J, Isaacs W, Carpten JD, Trent JM, Schleutker J, Barrett JC, Jurka J, Larionov V, Dynamic structure of the SPANX gene cluster mapped to the prostate cancer susceptibility locus HPCX at Xq27 Genome research15:1477-86 2005

PubMed ID: 16251457

Hirschhorn R, Identification of two new missense mutations (R156C and S291L) in two ADA- SCID patients unusual for response to therapy with partial exchange transfusions. Hum Mutat1:166-8 1992

PubMed ID: 1284479

Hirschhorn R, Ellenbogen A, Tzall SHirschhorn, Five missense mutations at the adenosine deaminase locus (ADA) detected by altered restriction fragments and their frequency in ADA--patients with severe combined immunodeficiency (ADA-SCID). Am J Hum Genet42:201-7 1992

PubMed ID: 1346349

Hirschhorn R, Chakravarti V, Puck J, Douglas SD, Homozygosity for a newly identified missense mutation in a patient with very severe combined immunodeficiency due to adenosine deaminase deficiency (ADA-SCID). Am J Hum Genet49:878-85 1991

PubMed ID: 1680289

Markert ML, Norby-Slycord C, Ward FE, A high proportion of ADA point mutations associated with a specific alanine-to-valine substitution. Am J Hum Genet45:354-61 1989

PubMed ID: 2773932

Tzall S, Ellenbogen A, Eng F, Hirschhorn R, Identification and characterization of nine RFLPs at the adenosine deaminase (ADA) locus. Am J Hum Genet44:864-75 1989

PubMed ID: 2567118

Akeson AL, Wiginton DA, Dusing MR, States JC, Hutton JJ, Mutant human adenosine deaminase alleles and their expression by transfection into fibroblasts. J Biol Chem263:16291-6 1988

PubMed ID: 3182793

Akeson AL, Wiginton DA, States JC, Perme CM, Dusing MR, Hutton JJ, Mutations in the human adenosine deaminase gene that affect protein structure and RNA splicing. Proc Natl Acad Sci U S A84:5947-51 1987

PubMed ID: 3475710

Berkvens TM, Gerritsen EJ, Oldenburg M, Breukel C, Wijnen JT, van Ormondt H, Vossen JM, van der Eb AJ, Meera Khan P, Severe combined immune deficiency due to a homozygous 3.2-kb deletion spanning the promoter and first exon of the adenosine deaminase gene. Nucleic Acids Res15:9365-78 1987

PubMed ID: 3684597

Danton MJ, Coleman MS, Isolation of mutant adenosine deaminase by coformycin affinity chromatography. Anal Biochem159:233-9 1986

PubMed ID: 3492941

Kantoff PW, Kohn DB, Mitsuya H, Armentano D, Sieberg M, Zwiebel JA, Eglitis MA, McLachlin JR, Wiginton DA, Hutton JJ, et al, Correction of adenosine deaminase deficiency in cultured human T and B cells by retrovirus-mediated gene transfer. Proc Natl Acad Sci U S A83:6563-7 1986

PubMed ID: 3489233

Valerio D, Dekker BM, Duyvesteyn MG, van der Voorn L, Berkvens TM, van Ormondt H, van der Eb AJ, One adenosine deaminase allele in a patient with severe combined immunodeficiency contains a point mutation abolishing enzyme activity. EMBO J5:113-9 1986

PubMed ID: 3007108

Adrian GS, Wiginton DA, Hutton JJ, Characterization of normal and mutant adenosine deaminase messenger RNAs by translation and hybridization to a cDNA probe. Hum Genet68:169-72 1984

PubMed ID: 6548726

Adrian GS, Wiginton DA, Hutton JJ, Structure of adenosine deaminase mRNAs from normal and adenosine deaminase-deficient human cell lines. Mol Cell Biol4:1712-7 1984

PubMed ID: 6208479

Daddona PE, Shewach DS, Kelley WN, Argos P, Markham AF, Orkin SH, Human adenosine deaminase. cDNA and complete primary amino acid sequence. J Biol Chem259:12101-6 1984

PubMed ID: 6090454

Valerio D, Duyvesteyn MG, van Ormondt H, Meera Khan P, van der Eb AJ, Adenosine deaminase (ADA) deficiency in cells derived from humans with severe combined immunodeficiency is due to an aberration of the ADA protein. Nucleic Acids Res12:1015-24 1984

PubMed ID: 6198631

Daddona PE, Mitchell BS, Meuwissen HJ, Davidson BL, Wilson JM, Koller CA, Adenosine deaminase deficiency with normal immune function. An acidic enzyme mutation. J Clin Invest72:483-92 1983

PubMed ID: 6603477

Wiginton DA, Hutton JJ, Immunoreactive protein in adenosine deaminase deficient human lymphoblast cell lines. J Biol Chem257:3211-7 1982

PubMed ID: 6977542

Daddona PE, Kelley WN, Characteristics of an aminohydrolase distinct from adenosine deaminase in cultured human lymphoblasts. Biochim Biophys Acta658:280-90 1981

PubMed ID: 6972784

Hirschhorn R, Roegner V, Jenkins T, Seaman C, Piomelli S, Borkowsky W, Erythrocyte adenosine deaminase deficiency without immunodeficiency. Evidence for an unstable mutant enzyme. J Clin Invest64:1130-9 1979

PubMed ID: 479373