Structural and functional basis of the developmental regulation of human coagulation factor IX gene: factor IX Leyden.

Hirosawa, Shinsaku; Fahner, James B.; Salier, Jean‐Philippe; Wu, Chin-Tuan Brocade; Lovrien, Evert W.; Kurachi, Kotoku

doi:10.1073/pnas.87.12.4421

Cited by 46 publications

(46 citation statements)

References 25 publications

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“…All of those studies were performed in vitro, with the implicit assumption that the puberty-related male sex hormone surge is directly involved. In contrast, our earlier works in vitro did not support a direct role for either testosterone or androgen receptors (13,14). One transgenic mouse model study was previously reported for a hemophilia B Leyden mutation, A ϩ 13G (20).…”

contrasting

confidence: 39%

“…1A, all of these mutations are clustered in an approximately 50 base pair (bp) region, nucleotide (nt) -34 through nt ϩ19 (see ref. 12 for nt numbering), known as the Leyden-specific region (LSR) in the 5Ј UTR (13,14). The wild-type gene LSR is known to bind 3 nuclear proteins, HNF-4 (15), an unidentified protein binding to the region spanning the nt -6 site (hereafter called UKP-6) (13) and C/EBP (16).…”

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confidence: 99%

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An age-related homeostasis mechanism is essential for spontaneous amelioration of hemophilia B Leyden

Kurachi

Huo

Ameri

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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Regulation of age-related changes in gene expression underlies many diseases. We previously discovered the first puberty-onset gene switch, the age-related stability element (ASE)/age-related increase element (AIE)-mediated genetic mechanism for agerelated gene regulation. Here, we report that this mechanism underlies the mysterious puberty-onset amelioration of abnormal bleeding seen in hemophilia B Leyden. Transgenic mice robustly mimicking the Leyden phenotype were constructed. Analysis of these animals indicated that ASE plays a central role in the puberty-onset amelioration of the disease. Human factor IX expression in these animals was reproducibly nullified by hypophysectomy, but nearly fully restored by administration of growth hormone, being consistent with the observed sex-independent recovery of factor IX expression. Ets1 was identified as the specific liver nuclear protein binding only to the functional ASE, G/CAG-GAAG, and not to other Ets consensus elements. This study demonstrates the clinical relevance of the first discovered pubertyonset gene switch, the ASE/AIE-mediated regulatory mechanism.factor IX ͉ growth hormone ͉ sex hormone ͉ puberty ͉ gene switch

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confidence: 39%

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confidence: 99%

An age-related homeostasis mechanism is essential for spontaneous amelioration of hemophilia B Leyden

Kurachi

Huo

Ameri

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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“…It has been found that promoter mutations in factor IX (4,7,12,19,28,39,45,47,49), protein C (2), haptoglobin (16), and ␤-globin (10,31,(36)(37)(38) lead to disruption of the binding of transcription factors, with a resulting clinically significant reduction of gene expression. Such a lesion might be expected to affect the level of expression of the factor VIII protein and give rise to a clinical phenotype.…”

Section: Discussionmentioning

confidence: 99%

Role of the Liver-Enriched Transcription Factor Hepatocyte Nuclear Factor 1 in Transcriptional Regulation of the Factor VIII Gene

McGlynn

Mueller

Begbie

et al. 1996

Molecular and Cellular Biology

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Coagulation factor VIII is an essential cofactor required for normal hemostatic function. A deficiency in factor VIII results in the bleeding disorder hemophilia A. Despite the fact that the factor VIII gene was cloned a decade ago, the mechanisms which control its transcription remain unresolved. In our studies, we have characterized 12 protein binding sites within the factor VIII promoter by DNase I protection assays performed with rat liver nuclear extracts. Three of these elements (sites 1 to 3) are situated within the 5 untranslated region of the gene, while three other sites (sites 4 to 6) lie within the first 100 bp upstream of the transcriptional start site. We have identified an additional site (site 7) ϳ300 bp upstream from site 6, as well as a cluster of five sites in a 250-bp region which terminates ϳ1 kb from the transcriptional start site. Seven of these binding sites (sites 2, 3, 4, 6, 7, 9, and 10) bind members of the C/EBP family of transcription factors. DBP also binds to five of these sites (sites 3, 4, 6, 7, and 9). Utilizing transient transfection studies in HepG2 cells, we have shown that deletion of the factor VIII promoter sequences distal to nucleotide ؊44 results in a significant but small increase in promoter activity. The activity of each of the various 5 deletion constructs is significantly enhanced by cotransfection of C/EBP␣ and D-site-binding protein expression plasmids, while cotransfection of both C/EBP␣ and C/EBP␤ plasmids resulted in a further enhancement of transactivation. These studies also provide evidence of a repressor element located between nucleotides ؊740 and ؊1002. Since the minimal promoter sequence (؊44 to ؉148) maintains the transcriptional activity of the full-length promoter sequence, we proceeded to identify additional factors binding to sites 1 to 4. Competition studies revealed that a ubiquitous transcription factor, NF-Y, binds to site 4, while the liver-enriched transcription factor hepatocyte nuclear factor I (HNF-1) binds to site 1. Mutation analysis of the minimal promoter demonstrated that HNF-1 is critical for activating transcription of the factor VIII gene in vitro. Our results also suggest that the multiple upstream elements that we have identified may act as a backup regulatory region in the event of disruption of the HNF-1 element in the 5 untranslated region.

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“…The amphotropic retroviruses of dLSNBAIX and dLSNBTIX were prepared by transfecting vector DNA into packaging cells, SCRIP (19), with the calcium phosphate precipitation method (20). The LIXSN retrovirus was prepared by transfecting vector DNA, LIXSN, into the ecotropic packaging cell line, (-2 (21) Proc.…”

Section: Kb) a Bamhi Fragment Containingmentioning

confidence: 99%

“…Clotting Activity of Factor IX. Factor IX activity was determined by the one-stage clotting assay employing human factor IX-deficient plasma (20). Normal human plasma pooled from 30 healthy individuals who showed normal blood clotting parameters was used as a standard for ELISA and the clotting assay.…”

Section: Kb) a Bamhi Fragment Containingmentioning

confidence: 99%

Expression of human factor IX in rat capillary endothelial cells: toward somatic gene therapy for hemophilia B.

Yao

Wilson

Nabel

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

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In aiming to develop a gene therapy approach for hemophilia B, we expressed and characterized human factor IX in rat capillary endothelial cells (CECs). Moloney murine leukemia virus-derived retrovirus vectors that contain human factor IX cDNA linked to heterologous promoters and the neomycin-resistant gene were constructed and employed to prepare recombinant retroviruses. Rat CECs and NIH 3T3 cells infected with these viruses were selected with the neomycin analogue, G418 sulfate, and tested for expression of factor IX. A construct with the factor IX cDNA under direct control by long terminal repeat gave the highest level of expression (0.84 and 3.6 pg per 106 cells per day for CECs and NIH 3T3 cells, respectively) as quantitated by immunoassays as well as clotting activity assays. A single RNA transcript of 4.4 kilobases predicted by the construct and a recombinant factor IX of 68 kilodaltons identical to purified plasma factor IX were found. The recombinant human factor IX produced showed full clotting activity, demonstrating that CECs have an efficient mechanism for posttranslational modifications, including y-carboxylation, essential for its biological activity. These results, in addition to other properties of the endothelium, including large number ofcells, accessibility, and direct contact with the circulating blood, suggest that CECs can serve as an efficient drug delivery vehicle producing factor IX in a somatic gene therapy for hemophilia B.

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Structural and functional basis of the developmental regulation of human coagulation factor IX gene: factor IX Leyden.

Cited by 46 publications

References 25 publications

An age-related homeostasis mechanism is essential for spontaneous amelioration of hemophilia B Leyden

An age-related homeostasis mechanism is essential for spontaneous amelioration of hemophilia B Leyden

Role of the Liver-Enriched Transcription Factor Hepatocyte Nuclear Factor 1 in Transcriptional Regulation of the Factor VIII Gene

Expression of human factor IX in rat capillary endothelial cells: toward somatic gene therapy for hemophilia B.

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