Identification of a novel promoter mutation in the human pyruvate kinase (<i>PK</i>) <i>LR</i> gene of a patient with severe haemolytic anaemia

Kugler, Wilfried; Laspe, Petra; Stahl, Markus; Schröter, W.; Lakomek, M.

doi:10.1046/j.1365-2141.1999.01386.x

Cited by 12 publications

(7 citation statements)

References 14 publications

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“…20,25,26 Initially, a Ϫ249delA mutation in the PK-R promoter was reported in association with PK deficiency. 27 However, reinvestigation of the patient's DNA revealed the presence of the same 3 in cis mutations we report here. 27 Kugler has allowed us to examine a DNA sample from his patient, and we have confirmed his results (data not shown).…”

Section: Discussionmentioning

confidence: 77%

Disruption of a novel regulatory element in the erythroid-specific promoter of the human PKLR gene causes severe pyruvate kinase deficiency

Wijk

Solinge

Nerlov

et al. 2003

Blood

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We established the molecular basis for pyruvate kinase (PK) deficiency in a white male patient with severe nonspherocytic hemolytic anemia. The paternal allele exhibited the common PKLR cDNA sequence (c.) 1529G>A mutation, known to be associated with PK deficiency. On the maternal allele, 3 in cis mutations were identified in the erythroid-specific promoter region of the gene: one deletion of thymine ؊248 and 2 single nucleotide substitutions, nucleotide (nt) ؊324T>A and nt ؊83G>C. Analysis of the patient's RNA demonstrated the presence of only the 1529A allele, indicating severely reduced transcription from the allele linked to the mutated promoter region. Transfection of promoter constructs into erythroleukemic K562 cells showed that the most upstream ؊324T>A and ؊248delT mutations were nonfunctional polymorphisms. In contrast, the ؊83G>C mutation strongly reduced promoter activity. Sitedirected mutagenesis of the promoter region revealed the presence of a putative regulatory element (PKR-RE1) whose core binding motif, CTCTG, is located between nt ؊87 and nt ؊83. Electrophoretic mobility shift assay using K562 nuclear extracts indicated binding of an as-yetunidentified trans-acting factor. This novel element mediates the effects of factors necessary for regulation of pyruvate kinase gene expression during red cell differentiation and maturation. (Blood.

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Section: Discussionmentioning

confidence: 77%

Disruption of a novel regulatory element in the erythroid-specific promoter of the human PKLR gene causes severe pyruvate kinase deficiency

Wijk

Solinge

Nerlov

et al. 2003

Blood

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“…They are spread all over the coding region, with no preference for the active site as determined by crystallographic analysis of the cat muscle enzyme (Muirhead et al , 1986; Allen & Muirhead, 1996). So far, only three mutations have been reported in the promoter region (van Solinge et al , 1997; Kugler et al , 1999). The mutations identified are mostly missense, splicing and stop codon, rarely small deletions, insertions and frameshift mutations.…”

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confidence: 99%

Molecular characterization of the PK‐LR gene in sixteen pyruvate kinase‐deficient patients

et al. 2001

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We studied the PK‐LR gene in 16 unrelated patients with congenital haemolytic anaemia associated with erythrocyte pyruvate kinase deficiency. Fifteen different mutations were detected among the 28 mutated alleles identified: two deletions (del 1010G, del 1042–1044); one four nucleotide duplication (nt 1515–1518, GGTC); one splice site [IVS6(−2)t]; nine missense (991A, 1003A, 1151T, 1160G, 1181T, 1181A, 1456T, 1483A, 1529A); and two nonsense (721T, 1675T) mutations. Eight of them [del 1010G, del 1042–1044, dupl 1515–1518, IVS6(−2)t, 1003A, 1160G, 1181T, 1181A] were novel. The deletion 1042–1044 causes the loss of Lys 348. Deletion 1010G and duplication 1515–1518 determine a frameshift and the creation of a stop codon at nucleotides 1019 and 1554 respectively. Mutation IVS6(−2)t leads to an alteration of the 5′ and 3′ splice site consensus sequence; the cDNA analysis shows a 67‐bp deletion in the first part of exon 11 (del 1437–1503). All the four new missense mutations involve highly conserved amino acids. The most frequent mutation in Italy would appear to be 1456T. Correlation was made between mutations, biochemical characteristics of the enzyme and clinical course of the disease.

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“…Mutations in the PKLR gene affecting the R‐type isoenzyme, exclusively expressed in erythrocytes, are responsible for the PK deficiency ( Beutler & Baronciani, 1996; Miwa & Fujii, 1996). Most of the described mutations are in the coding regions: 11 are splice‐site mutations and, recently, three mutations were found in the promoter region of the PKLR gene ( Baronciani et al , 1998 ; Kugler et al , 1999 ).…”

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confidence: 99%

A new PKLR gene mutation in the R‐type promoter region affects the gene transcription causing pyruvate kinase deficiency

Manco

Ribeiro²,

Máximo³

et al. 2000

Br J Haematol

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Summary. Mutations in the PKLR gene responsible for pyruvate kinase (PK)-deficient anaemia are mainly located in the coding regions: 11 are in the splicing sites and, recently, three mutations have been described in the promoter region. We now report a novel point mutation A3G on nucleotide 72, upstream from the initiation codon of the PKLR gene, in four Portuguese PK-deficient patients. This new regulatory mutation occurs within the most proximal of the four GATA motifs (GATA-A element) in the R-type promoter region. In two patients who were homozygous for this mutation, a semiquantitative reverse transcription polymerase chain reaction (PCR) procedure was used to evaluate the amount of R-PK mRNA transcript in the reticulocytes. The mRNA level was about five times lower than in normal controls, demonstrating that the PKLR gene transcription is severely affected, most probably because the 272A3G point mutation disables the binding of the erythroid transcription factor GATA-1 to the GATA-A element. Supporting these data, the two patients homozygous for the 272A3G mutation had severe haemolytic anaemia and were transfusion dependent until splenectomy. Two other patients who were compound heterozygous for this mutation and the previously described missense mutation 1456C3T had a mild condition.

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Identification of a novel promoter mutation in the human pyruvate kinase (PK) LR gene of a patient with severe haemolytic anaemia

Cited by 12 publications

References 14 publications

Disruption of a novel regulatory element in the erythroid-specific promoter of the human PKLR gene causes severe pyruvate kinase deficiency

Disruption of a novel regulatory element in the erythroid-specific promoter of the human PKLR gene causes severe pyruvate kinase deficiency

Molecular characterization of the PK‐LR gene in sixteen pyruvate kinase‐deficient patients

A new PKLR gene mutation in the R‐type promoter region affects the gene transcription causing pyruvate kinase deficiency

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