GM02606

GM02606 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:

Female

Age:

8 MO (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

White

Country of Origin

USA

Family Member

Relation to Proband

proband

Confirmation

Clinical summary/Case history

Species

Homo sapiens

Common Name

Human

Remarks

Clinically affected; deficient ADA activity in RBC's, lymphoid line, and fibroblasts; enzyme phenotypes: 6PGD=A, Pepitdases A,C, and D= 1; Acid A-glucosidase=1, Neutral A-glucosidase C=1; < 1% of normal ADA activity and CRM protein in lymph culture; normal ADA mRNA; donor subject is a compound heterozygote: one allele has a C>T transition at nucleotide 396 in exon 4 of the ADA gene [396C>T] resulting in a substitution of tryptophan for arginine at codon 101 [Arg101Trp(R101W)] and a second 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)]; same donor as GM02605 fibroblast; mother is GM02607(fibro)/GM02608 (lymph) and father is GM02609(fibro)/GM02610(lymph); clinical documentation and in-house testing via amelogenin and DYS227 PCR assays have identified this subject as female.

Characterizations

IDENTIFICATION OF SPECIES OF ORIGIN

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

MUTATION VERIFICATION

Adrian et al (Mol Cell Biol 4:1712-1717 1984) performed S1 endonuclease cleavage of ADA cDNA-mRNA hybrids to show that the ADA mRNA from this culture yielded fragments that were indistinguishable from ADA mRNA fragments from normal controls. The ADA cDNA utilized for these experiments represented the complete mRNA sequence with the possible exception of some of the 5 prime untranslated region. Akeson et al (J Biol Chem 263:16291-16296 1988) reported the sequence analysis of ADA cDNA from this ADA-deficient cell line. One allele has a mutation altering amino acid 101 from Arg to Trp (base 396 in exon 4 is T rather than C). The other allele has a mutation altering amino acid 211 from Arg to His (base 727 in exon 7 is A rather than G). Insertion of the mutant ADA cDNA into an expression vector and testing in a transient expression system revealed that 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 same mutation in amino acid no. 211 (Arg to His) had also been observed for another ADA-deficient cell line (GM02756).

adenosine deaminase

According to the submitter, biochemical test results for this subject showed decreased enzyme activity. EC Number: 3.5.4.4

Gene

ADA

Chromosomal Location

20q13.11

Allelic Variant 1

608958.0002; SEVERE COMBINED IMMUNODEFICIENCY, AUTOSOMAL RECESSIVE, T CELL-NEGATIVE, B CELL-NEGATIVE, NK CELL-NEGATIVE, DUE TO ADENOSINE DEAMINASE DEFICIENCY

Identified Mutation

ARG101TRP; Akeson et al. [J Biol Chem 263:16291 (1988)] summarized the point mutations identified in ADA deficiency cases. They came from 5 different patients, each of whom proved to be a compound heterozygote. GM02606 was found to have change of arg101 to trp resulting from a change of CGG to TGG as well as substitution of his for arg211 (102700.0004) as a result of change of CGT to CAT.

Gene

ADA

Chromosomal Location

20q13.11

Allelic Variant 2

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.

Phenotypic Data

Remarks

Publications

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

Akeson AL, Wiginton DA, Hutton JJ, Normal and mutant human adenosine deaminase genes Journal of cellular biochemistry39:217-28 1989

PubMed ID: 2651461

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

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

Adrian GS, Hutton JJ, Adenosine deaminase messenger RNAs in lymphoblast cell lines derived from leukemic patients and patients with hereditary adenosine deaminase deficiency. J Clin Invest71:1649-60 1983

PubMed ID: 6134754

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, Adrian GS, Friedman RL, Suttle DP, Hutton JJ, Cloning of cDNA sequences of human adenosine deaminase. Proc Natl Acad Sci U S A80:7481-5 1983

PubMed ID: 6200875

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