An Sp1 binding site mutation of the <i>PROS1</i> promoter in a patient with protein S deficiency

Sanda, Naomi; Fujimori, Yuta; Kashiwagi, Takahiro; Takagi, Akira; Murate, Takashi; Mizutani, Emi; Matsushita, Tadashi; Naoe, Tomoki; Kojima, Tetsuhito

doi:10.1111/j.1365-2141.2007.06668.x

Cited by 14 publications

(8 citation statements)

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“…Therefore, the molecular basis of PSD is clearly heterogeneous. It is interesting that the previously reported mutations [42][43][44][45] identified in this study were mainly from Japanese, indicating that East Asian populations may share a similar genetic background. However, ProS Tokushima (p.K196E), which is a prevalent ProS variant in the Japanese ProS-deficient patients and was identified in 6-9% of venous thrombosis patients [49], was not observed in the cohort of the present study.…”

Section: Discussionmentioning

confidence: 89%

Molecular basis of protein S deficiency in China

et al. 2013

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Protein S (ProS) is a physiological inhibitor of coagulation with an important function in the downregulation of thrombin generation. ProS deficiency is a major risk factor for venous thrombosis. This study enrolled 40 ProS-deficient probands to investigate the molecular basis of hereditary ProS deficiency in Chinese patients. A mutation analysis was performed by resequencing the PROS1 gene. Large deletions were identified by multiplex ligation-dependent probe amplification (MLPA) analysis. A total of 20 different mutations, including 15 novel mutations, were identified in 21 of the 40 index probands. Small mutations were detected in 18 (45.0%) probands, and large deletions were found in 3 (7.5%) probands, leaving 19 (47.5%) patients without causative variants. To evaluate the functional consequences of 2 novel missense variants, ex vivo thrombin-generation assays, bioinformatics tools, and in vitro expression studies were employed. The p.Asn365Lys ProS variant was found to have moderately impaired secretion and reduced activated protein C cofactor activity. In contrast, the p.Pro410His mutant appeared to have severely impaired secretion but full anticoagulant activity. This study is the largest investigation of ProS deficiency in China and the first investigation of the influence of Type I ProS missense mutations on the global level of coagulation function. The p.K196E mutation, which is common in the neighboring Japanese population, was not found in our Chinese population, and null mutations were common in our Chinese population but not common in Japan. Further genetic analysis is warranted to understand the causes of ProS deficiency in patients without a genetic explanation. Am. J.

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Section: Discussionmentioning

confidence: 89%

Molecular basis of protein S deficiency in China

et al. 2013

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“…These data suggest that the distal GC-rich motif may be more important than the proximal GC-rich motif. A patient carrying the g.Ϫ168cϾt mutation showed decreased levels of plasma PS but supposedly did not show E 2 -dependent PROS1 repression (53).…”

Section: Discussionmentioning

confidence: 95%

“…VEGFR2 is also regulated both positively and negatively by ER␣-Sp protein interaction, but the type of regulation depends on the cell line, MCF7 or ZR-75 (51,52). Interestingly, we reported previously a novel missense mutation at Ϫ168 from the ATG of the PROS1 promoter (g.Ϫ168cϾt), which results in weak promoter activity of PROS1 (53). The mutation is located in the distal GC-rich motif, which could be critical for E 2 -dependent repression, as shown in the current study.…”

Section: Discussionmentioning

confidence: 98%

Down-regulation of PROS1 Gene Expression by 17β-Estradiol via Estrogen Receptor α (ERα)-Sp1 Interaction Recruiting Receptor-interacting Protein 140 and the Corepressor-HDAC3 Complex

Suzuki

Sanda

Miyawaki

et al. 2010

Journal of Biological Chemistry

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Pregnant women show a low level of protein S (PS) in plasma,which is known to be a risk for deep venous thrombosis. 17␤-Estradiol (E 2 ), an estrogen that increases in concentration in the late stages of pregnancy, regulates the expression of various genes via the estrogen receptor (ER). Here, we investigated the molecular mechanisms behind the reduction in PS levels caused by E 2 in HepG2-ER␣ cells, which stably express ER␣, and also the genomic ER signaling pathway, which modulates the liganddependent repression of the PS␣ gene (PROS1). We observed that E 2 repressed the production of mRNA and antigen of PS. A luciferase reporter assay revealed that E 2 down-regulated PROS1 promoter activity and that this E 2 -dependent repression disappeared upon the deletion or mutation of two adjacent GCrich motifs in the promoter. An electrophoretic mobility shift assay and DNA pulldown assay revealed that the GC-rich motifs were associated with Sp1, Sp3, and ER␣. In a chromatin immunoprecipitation assay, we found ER␣-Sp protein-promoter interaction involved in the E 2 -dependent repression of PROS1 transcription. Furthermore, we demonstrated that E 2 treatment recruited RIP140 and the NCoR-SMRT-HDAC3 complex to the PROS1 promoter, which hypoacetylated chromatin. Taken together, this suggested that E 2 might repress PROS1 transcription depending upon ER␣-Sp1 recruiting transcriptional repressors in HepG2-ER␣ cells and, consequently, that high levels of E 2 leading to reduced levels of plasma PS would be a risk for deep venous thrombosis in pregnant women.

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“…A second mutation was identified in the 5' flanking sequence of the PROS1 gene ( Figure 1B), and represented a heterozygous CG nucleotide substitution at a position 190 bp upstream of the translational start site ( Figure 1C). This substitution (PROS1 g.-190 C>G) is present within a proposed Sp1 transcription factor binding site that is highly conserved among mammals [21].…”

Section: Resultsmentioning

confidence: 99%

“…However, the G67A mutation alone is sufficient to result in the PS deficiency phenotype presented by the propositus [19]. Although another mutation has been identified in the PROS1 promoter region in a PS deficient patient [21], the current mutation has not been previously reported in the ISTH PROS1 mutation database [10]. Further studies would be required to confirm that the mutated sequence reduces the promoter activity in the mutant PROS1 gene but a reduction in PROS1 gene transcription with a resulting reduction in PS mRNA and plasma protein is consistent with the patient's hematological presentation.…”

Section: Discussionmentioning

confidence: 99%

Identification of two novel mutations associated with combined protein C and protein S deficiency

Smith¹,

Carter²,

Smith³

et al. 2017

Vascul Dis Ther

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We have studied a 60 year old male of Chinese descent who presented with combined deficiency of the anticoagulant proteins C and S. Molecular genetic analysis of the patient's PROC gene revealed a novel heterozygous mutation that resulted in a V221G mutation. Two mutations were identified in the patient's PROS1 gene:(1) an E67E mutation that has been reported previously and is associated with severe protein S deficiency, and (2) a novel mutation in the 5' flanking sequence that changes a putative binding site for the transcription factor Sp 1. Together, these mutations are consistent with the combined protein C and S deficiency symptoms presented by the patient.

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An Sp1 binding site mutation of the PROS1 promoter in a patient with protein S deficiency

Cited by 14 publications

References 10 publications

Molecular basis of protein S deficiency in China

Molecular basis of protein S deficiency in China

Down-regulation of PROS1 Gene Expression by 17β-Estradiol via Estrogen Receptor α (ERα)-Sp1 Interaction Recruiting Receptor-interacting Protein 140 and the Corepressor-HDAC3 Complex

Identification of two novel mutations associated with combined protein C and protein S deficiency

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