The G20210A mutation does not affect the stability of prothrombin mRNA in vivo

Pollak, Eleanor S.; Lam, Hieu; Russell, Janice

doi:10.1182/blood-2002-02-0412

Cited by 47 publications

(42 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…However, based on the knowledge that the G20210A substitution represents in vitro a gain-of-function mutation causing mRNA accumulation and increased protein synthesis (7 ), creates in vitro but not in vivo a more effective polyadenylation site and cleavage site (7)(8)(9), and is proposed to be associated with different mRNA structures leading to abnormal mRNA function (9 ), it will be worthwhile to initiate additional studies to estimate the impact of this genetic variant on risk assessment for thrombotic events and adverse pregnancy outcomes. Improved Method for Isolating CellFree DNA…”

Section: Atypical Melting Curve Resulting From Genetic Variation In Tmentioning

confidence: 99%

Atypical Melting Curve Resulting from Genetic Variation in the 3′ Untranslated Region at Position 20218 in the Prothrombin Gene Analyzed with the LightCycler Factor II (Prothrombin) G20210A Assay

et al. 2005

View full text Add to dashboard Cite

Section: Atypical Melting Curve Resulting From Genetic Variation In Tmentioning

confidence: 99%

Atypical Melting Curve Resulting from Genetic Variation in the 3′ Untranslated Region at Position 20218 in the Prothrombin Gene Analyzed with the LightCycler Factor II (Prothrombin) G20210A Assay

et al. 2005

View full text Add to dashboard Cite

Section: Introductionmentioning

confidence: 92%

“…Pollak et al described that in the case of the 20210A mutation 100% of transcripts were polyadenylated at position 20210, while in the case of the wild-type 20210G cleavage site with unknown 19911 genotype most (74%) of the transcripts were polyadenylated at position 20212 and only 26% at position 20210. 17 It is noteworthy that for 20210G, polyadenylation has most likely occurred at position 20211. If the start of the poly-A tail corresponds with an A in the genomic sequence, this is the base to which the tail is added after transcription.…”

Section: Cleavage Site Heterogeneity Depending On Base 20210mentioning

confidence: 99%

“…A different study provided no evidence in vivo for altered prothrombin mRNA stability by the 20210A mutation. 17 In contrast, the in vitro study by Carter et al 18 found an effect of the 20210A genotype on both the efficiency of mRNA processing and mRNA stability.In addition to these contradictory findings, there is evidence 19,20 that a common single nucleotide polymorphism (SNP) (19911AϾG) 21 in intron M, the prothrombin gene's last intron, may have a functional role. A gene dosage effect of the 19911G allele on prothrombin activity was shown in vivo.…”

mentioning

confidence: 92%

See 1 more Smart Citation

The intronic prothrombin 19911A>G polymorphism influences splicing efficiency and modulates effects of the 20210G>A polymorphism on mRNA amount and expression in a stable reporter gene assay system

Ahsen

Oellerich

2004

Blood

View full text Add to dashboard Cite

The common prothrombin gene cleavage site mutation 20210G>A is associated with elevated prothrombin levels and thrombosis. The pathomechanism of the 20210G>A mutation was explained by increased mRNA formation and/or more efficient translation. Human studies also showed an influence of the intronic 19911A>G polymorphism on prothrombin activity. We established HepG2 cell lines stably transfected with prothrombin mini-genes containing the last 2 prothrombin exons, the last intron, 3 untranslated region (UTR), and flanking sequence. The highest mRNA expression and protein activity resulted from the mutant haplotype 19911A-20210A. Haplotypes with wild-type cleavage site (19911A-20210G, 19911G-20210G) also differed significantly as a consequence of the intronic 19911 mutation; the 19911G-20210G haplotype showed lower expression than the 19911A-20210G haplotype, whereas previous clinical studies have reported elevated prothrombin activity with the 19911G-20210G haplotype. The cleavage site pattern was homogeneous with 20210A, which may cause a favorable intracellular processing, and heterogeneous with 20210G. In an independent assay for splicing efficiency, 19911G showed about 30% higher efficiency than 19911A. We conclude that the intronic 19911A>G single nucleotide polymorphism is itself functional and changes splicing efficiency by altering a known functional pentamer motif. Further studies are needed to define the value of additional prothrombin 19911 genotyping for thrombophilia screening, especially in cases heterozygous for 20210G>A. IntroductionA noncoding mutation in the 3Ј untranslated region (UTR) of the prothrombin gene (20210GϾA) has been associated with thrombophilia. 1 Several studies have since confirmed the association of this common mutation with arterial 2 or venous 1,3-7 thrombosis. The 20210GϾA mutation confers a 3-to 7-fold increased risk for venous thrombosis. 8,9 The determination of the prothrombin 20210GϾA genotype is recommended as a high-priority test in the investigation of genetic thrombophilia. 10 The activity of prothrombin may be partially regulated at a posttranslational level 11 and is controlled by a strong genetic background. 12 Linkage analysis demonstrated a cosegregation of the mutant allele with elevated prothrombin activity and suggested that this mutation in the 3Ј UTR is in itself functional instead of in linkage disequilibrium with another polymorphism. 13 Recent reviews have emphasized the role of the mRNA UTR in the pathomechanisms of some human diseases. 14,15 Gehring et al demonstrated increased prothrombin mRNA end formation efficiency as a cause of elevated prothrombin protein levels in carriers of the 20210GϾA mutation. 16 In this study, a ␤-globin gene vector supplemented with the UTR of the prothrombin gene was used to investigate the polyadenylation reaction. A different study provided no evidence in vivo for altered prothrombin mRNA stability by the 20210A mutation. 17 In contrast, the in vitro study by Carter et al 18 found an effect of the 20210A genotype on bo...

show abstract

“…For ETP and peak, associations were observed with rs1799963 and rs3136516, 2 F2 variants already known to associate with thrombin generation and whose functionality has already been discussed. 20,[50][51][52][53][54] Both rare alleles, rs1799963-A and rs3136516-G, were associated independently from each other with increased thrombin generation, the strongest effect being at rs1799963. These 2 mutations act on thrombin generation through increase in prothrombin levels.…”

Section: Discussionmentioning

confidence: 99%

A meta-analysis of genome-wide association studies identifies ORM1 as a novel gene controlling thrombin generation potential

Rocañín-Arjó

Cohen

Carcaillon

et al. 2014

Blood

View full text Add to dashboard Cite

Key Points• Genetic variations at the ORM1 locus and concentrations of the encoded protein associate with thrombin generation.• These findings may guide the development of novel antithrombotic treatments.Thrombin, the major enzyme of the hemostatic system, is involved in biological processes associated with several human diseases. The capacity of a given individual to generate thrombin, called the thrombin generation potential (TGP), can be robustly measured in plasma and was shown to associate with thrombotic disorders. To investigate the genetic architecture underlying the interindividual TGP variability, we conducted a genome-wide association study in 2 discovery samples (N 5 1967) phenotyped for 3 TGP biomarkers, the endogenous thrombin potential, the peak height, and the lag time, and replicated the main findings in 2 independent studies (N 5 1254). We identified the ORM1 gene, coding for orosomucoid, as a novel locus associated with lag time variability, reflecting the initiation process of thrombin generation with a combined P value of P 5 7.1 3 10 215 for the lead single nucleotide polymorphism (SNP) (rs150611042). This SNP was also observed to associate with ORM1 expression in monocytes (P 5 8.7 3 10 210 ) and macrophages (P 5 3.2 3 10 23 ). In vitro functional experiments further demonstrated that supplementing normal plasma with increasing orosomucoid concentrations was associated with impaired thrombin generation. These results pave the way for novel mechanistic pathways and therapeutic perspectives in the etiology of thrombin-related disorders. (Blood. 2014;123(5):777-785)

show abstract

The G20210A mutation does not affect the stability of prothrombin mRNA in vivo

Cited by 47 publications

References 23 publications

Atypical Melting Curve Resulting from Genetic Variation in the 3′ Untranslated Region at Position 20218 in the Prothrombin Gene Analyzed with the LightCycler Factor II (Prothrombin) G20210A Assay

Atypical Melting Curve Resulting from Genetic Variation in the 3′ Untranslated Region at Position 20218 in the Prothrombin Gene Analyzed with the LightCycler Factor II (Prothrombin) G20210A Assay

The intronic prothrombin 19911A>G polymorphism influences splicing efficiency and modulates effects of the 20210G>A polymorphism on mRNA amount and expression in a stable reporter gene assay system

A meta-analysis of genome-wide association studies identifies ORM1 as a novel gene controlling thrombin generation potential

Contact Info

Product

Resources

About

The G20210A mutation does not affect the stability of prothrombin mRNA in vivo

Cited by 47 publications

References 23 publications

Atypical Melting Curve Resulting from Genetic Variation in the 3′ Untranslated Region at Position 20218 in the Prothrombin Gene Analyzed with the LightCycler Factor II (Prothrombin) G20210A Assay

Atypical Melting Curve Resulting from Genetic Variation in the 3′ Untranslated Region at Position 20218 in the Prothrombin Gene Analyzed with the LightCycler Factor II (Prothrombin) G20210A Assay

The intronic prothrombin 19911A&gt;G polymorphism influences splicing efficiency and modulates effects of the 20210G&gt;A polymorphism on mRNA amount and expression in a stable reporter gene assay system

A meta-analysis of genome-wide association studies identifies ORM1 as a novel gene controlling thrombin generation potential

Contact Info

Product

Resources

About

The intronic prothrombin 19911A>G polymorphism influences splicing efficiency and modulates effects of the 20210G>A polymorphism on mRNA amount and expression in a stable reporter gene assay system