Molecular comparison of delta beta-thalassemia and hereditary persistence of fetal hemoglobin DNAs: evidence of a regulatory area?

Ottolenghi, Sergio; Giglioni, Barbára; Taramelli, Roberto; Comi, Paola; Mazza, U; Saglio, Giuseppe; Camaschella, Clara; Izzo, Paola; Cao, Antonio; Galanello, Renzo; E, Gimferrer; Baiget, Montserrat; Gianni, AM

doi:10.1073/pnas.79.7.2347

Cited by 84 publications

(29 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The erythroid-specific hypersensitive site downstream of the HPFH-1 breakpoint is eliminated in Spanish 8513-thalassemia, which is characterized by smaller elevations of fetal hemoglobin in adult life than are HPFH-1 and -2 (10% in Spanish thalassemia versus 26 and 44% in HPFH-1 and -2, respectively [28]). The Spanish b°'-thalassemia deletion extends approximately 10 kb farther downstream than does HPFH-1 (6; Feingold and Forget, in press).…”

mentioning

confidence: 99%

“…4A). A strong correlation exists between the presence of DNase I-hypersensitive sites and binding sites for factors which regulate transcription (4,10,13,44 (28) suggests that deletion of intervening DNA alone is insufficient to account for the phenotype of deletional HPFH.…”

mentioning

confidence: 99%

“…However, no 3'-flanking regions which inhibit globin gene expression at a distance (and which would be eliminated by these deletions) have yet been described. Even relatively short deletions which eliminate the 8 and 1 genes display increased fetal hemoglobin in adult heterozygotes (6,28). One hypothesis is that trans-acting factors normally interact with the regulatory regions of the adult genes to activate adult gene transcription and inactivate fetal gene transcription.…”

mentioning

confidence: 99%

“…This phenotype is associated with the removal of sequences from the 3' end of the P-like globin gene locus, including the 8-and P-globin genes, thereby introducing DNA normally very far (-100 kb) 3' to the P-globin gene into the globin cluster (7). Shorter deletions which remove the 8 and p genes but end within the P-like gene cluster produce lower but significant amounts of fetal hemoglobin in adult life and are called (8p)0 thalassemias (28). By analogy to the (Byp) thalassemias, the mechanism of prolonged expression of the fetal globin genes could be either the elimination of sequences involved in the transcriptional shutoff of the y-globin genes during appropriate globin gene switching or the introduction of sequences capable of enhancing the expression of the adjacent y-globin genes on the deletion chromosome.…”

mentioning

confidence: 99%

“…1). This deletion is characterized by lower levels of fetal hemoglobin in adult life than is HPFH (28) serum and then subjected to elutriation as described previously (40). Fractions enriched for nucleated erythroid cells were pooled, suspended in medium, and used immediately.…”

mentioning

confidence: 99%

See 4 more Smart Citations

Translocation of an erythroid-specific hypersensitive site in deletion-type hereditary persistence of fetal hemoglobin.

Elder¹,

Forrester²,

Thompson³

et al. 1990

Mol. Cell. Biol.

View full text Add to dashboard Cite

Hereditary persistence of fetal hemoglobin (HPFH) can involve large deletions which eliminate the 3' end of the ,8-like globin gene cluster and more than 70 kilobases (kb) of flanking DNA. Blot hybridization revealed a DNase I-hypersensitive site extending from 1.1 to 1.4 kb downstream of the HPFH-1 3' deletion endpoint. The site was found in normal fetal and adult nucleated erythroid cells and in two erythroleukemia cell lines but not in nonerythroid cells and tissues. Simian virus 40 core enhancer-like sequences were found nonrandomly distributed within the boundaries of the site, which is contained in a fragment of known enhancer activity (E. A. Feingold and B. G. Forget, Blood, in press). A second hypersensitive site was found 0.5 kb upstream of the HPFH-1 3' deletion endpoint but was not erythroid specific. A third site, most prominent in fetal liver-derived erythroid cells, was found 1 kb upstream of the HPFH-2 deletion endpoint. As predicted by the locations of the deletion endpoints, the first two sites were translocated to within 12 kb of the A_Y gene in erythroid colonies derived from an HPFH-2 heterozygote and in hybrid mouse-human erythroid cells carrying the HPFH-2 deletion chromosome. Further analysis of this region showed that it was DNase I sensitive in erythroid and myeloid cells, indicating that it resides in an open chromatin domain. These observations suggest that alterations of chromatin structure flanking the fetal globin genes may contribute to abnormal gene regulation in deletion-type HPFH.

show abstract

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Translocation of an erythroid-specific hypersensitive site in deletion-type hereditary persistence of fetal hemoglobin.

Elder¹,

Forrester²,

Thompson³

et al. 1990

Mol. Cell. Biol.

View full text Add to dashboard Cite

show abstract

Evaluation of the BeTha Gene 1 Kit for the Qualitative Detection of the Eight Most Common Mediterranean β-Thalassemia Mutations

et al. 1998

View full text Add to dashboard Cite

We describe the evaluation of the Bio-Rad BeTha Gene 1 kit (Bio-Rad Laboratories, Hercules, CA), a DNA-probe assay designed for the qualitative determination of the eight most common Mediteranean-thalassemia mutations. The kit utilizes the principle of allele-specific oligonucleotide (ASO) hybridization. Following sample preparation and in vitro DNA amplification by the polymerase chain reaction (PCR), an allele-specific detection of the amplified products by a nonradioactive enzymatic assay is performed. Geno-mic DNA is prepared from an individual's whole blood with a DNA purification matrix. In a second step, the-globin gene is amplified in a multiplex PCR reaction containing four 5 biotinylated oligonucleotide primers. In a final step, an aliquot of the PCR reaction is first chemically denatured and then captured in two eight-well strips of a 96-well enzyme-linked immunosorbent assay (ELISA) plate by hybridization to an immobilized ASO probe. Each DNA sequence at each of the eight mutation sites is represented by one normal and one mutant ASO. During this capture/hybridization step, which is performed at 37°C, only perfectly matched PCR products will be captured by an ASO. Subsequently, the allele-specific captured biotin-labeled PCR products are detected by a colorimetric enzymatic reaction. The system permits the detection of 16-thalassemia alleles using a high-throughput format that can be automated easily. A clinical feasibility study was performed to evaluate the functionality (method camparison study, assay validity using samples previously collected and stored at various temperatures for different periods of time, interference on kit performance, and assay validity for prenatal diagnosis) and the usability (ease of use, sample throughput) of the kit. The analysis of 110 samples previously studied with reference methods showed 100% clinical sensitivity and specificity. We demonstrate here that the procedure not only increases the throughput of-thalassemia allele genotyping but also provides an accurate, rapid, reliable, and noniso-topic diagnostic tool. Am.

show abstract

Evaluation of the BeTha Gene 1 Kit for the Qualitative Detection of the Eight Most Common Mediterranean β‐Thalassemia Mutations

et al. 1998

View full text Add to dashboard Cite

We describe the evaluation of the Bio-Rad BeTha Gene 1 kit (Bio-Rad Laboratories, Hercules, CA), a DNA-probe assay designed for the qualitative determination of the eight most common Mediteranean ␤-thalassemia mutations. The kit utilizes the principle of allele-specific oligonucleotide (ASO) hybridization. Following sample preparation and in vitro DNA amplification by the polymerase chain reaction (PCR), an allele-specific detection of the amplified products by a nonradioactive enzymatic assay is performed. Genomic DNA is prepared from an individual's whole blood with a DNA purification matrix. In a second step, the ␤-globin gene is amplified in a multiplex PCR reaction containing four 5 biotinylated oligonucleotide primers. In a final step, an aliquot of the PCR reaction is first chemically denatured and then captured in two eight-well strips of a 96-well enzymelinked immunosorbent assay (ELISA) plate by hybridization to an immobilized ASO probe. Each DNA sequence at each of the eight mutation sites is represented by one normal and one mutant ASO. During this capture/hybridization step, which is performed at 37°C, only perfectly matched PCR products will be captured by an ASO. Subsequently, the allele-specific captured biotin-labeled PCR products are detected by a colorimetric enzymatic reaction. The system permits the detection of 16 ␤-thalassemia alleles using a high-throughput format that can be automated easily. A clinical feasibility study was performed to evaluate the functionality (method camparison study, assay validity using samples previously collected and stored at various temperatures for different periods of time, interference on kit performance, and assay validity for prenatal diagnosis) and the usability (ease of use, sample throughput) of the kit. The analysis of 110 samples previously studied with reference methods showed 100% clinical sensitivity and specificity. We demonstrate here that the procedure not only increases the throughput of ␤-thalassemia allele genotyping but also provides an accurate, rapid, reliable, and nonisotopic diagnostic tool. Am.

show abstract

Molecular comparison of delta beta-thalassemia and hereditary persistence of fetal hemoglobin DNAs: evidence of a regulatory area?

Cited by 84 publications

References 29 publications

Translocation of an erythroid-specific hypersensitive site in deletion-type hereditary persistence of fetal hemoglobin.

Translocation of an erythroid-specific hypersensitive site in deletion-type hereditary persistence of fetal hemoglobin.

Evaluation of the BeTha Gene 1 Kit for the Qualitative Detection of the Eight Most Common Mediterranean β-Thalassemia Mutations

Evaluation of the BeTha Gene 1 Kit for the Qualitative Detection of the Eight Most Common Mediterranean β‐Thalassemia Mutations

Contact Info

Product

Resources

About