Detection and characterization of seven novel protein S (PROS) gene lesions: evaluation of reverse transcript-polymerase chain reaction as a mutation screening strategy

Formstone, Caroline J.; Wacey, A. I.; Berg, Lutz-Peter; Rahman, Salman; Bevan, David; Rowley, Megan; Voke, Jennifer; Bernardi, Francesco; Legnani, Cristina; Simioni, Paolo; Girolami, Antonio; Tuddenham, E G D; Kakkar, Vijay V.; Cooper, David Neil

doi:10.1182/blood.v86.7.2632.2632

Cited by 37 publications

(25 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The 3 bp deletion in intron A that was identified in subject 7 has been described previously and was not associated with PS deficiency (Formstone et al , 1995). Our finding that the mRNA was expressed at similar levels from both PROS1 alleles in this subject would suggest that this mutation does not influence plasma PS levels and that the co‐inherited Heerlen allele was responsible for the reduction in levels observed in this case.…”

Section: Discussionsupporting

confidence: 57%

“…In addition to carrying the Heerlen allele, subject 7 was heterozygous for a 3 bp deletion from the sequence located 68–72 bp upstream of exon 2. This mutation has been identified previously and was not found to be associated with PS deficiency (Formstone et al , 1995). We examined the possibility that the mutation resulted in differential expression of the corresponding PROS1 mRNA using the heterozygous exonic Heerlen polymorphism as a marker of allele expression.…”

Section: Analysis Of Pros1 Gene Expressionmentioning

confidence: 64%

See 1 more Smart Citation

The prevalence of, and molecular defects underlying, inherited protein S deficiency in the general population

et al. 2004

View full text Add to dashboard Cite

SummaryThe molecular basis of protein S (PS) deficiency was investigated in seven of eight donors identified with persistently low plasma PS levels from a survey of PS levels in 3788 Scottish blood donors. PROS1 gene analysis identified at least one defect in six donors. Five were heterozygous for the Heerlen polymorphism predicting a Ser460Pro substitution. Haplotype analysis revealed the possibility that this allele was inherited with the same haplotype in four of the five donors, suggesting a founder effect for the Heerlen allele in this population. One Heerlen allele carrier was also heterozygous for a 3 bp deletion 68-72 bp upstream of exon 2. Platelet PROS1 transcript analysis showed no reduction in mRNA expression from the affected allele in this donor. A T to G transversion 3 bp upstream of exon 12 was identified in one donor, which is predicted to reduce the efficiency of PS mRNA splicing. However, PROS1 transcript analysis showed no evidence of exon skipping or cryptic splicing. No PROS1 gene defect was detected in the remaining donor. This genetic information enabled us to refine our estimate of the prevalence of heritable PS deficiency in the Scottish population to between 0AE16% and 0AE21%, predominantly resulting from the presence of the Heerlen allele.

show abstract

Section: Discussionsupporting

confidence: 57%

Section: Analysis Of Pros1 Gene Expressionmentioning

confidence: 64%

The prevalence of, and molecular defects underlying, inherited protein S deficiency in the general population

et al. 2004

View full text Add to dashboard Cite

show abstract

“…The identification of PROS1 mutations in PS deficiency has been reported in several studies, including more than 300 unrelated PS deficient families [10,17,23–33]. In 53% of all these cases a mutation, likely to cause the decreased level of PS, was found, leaving the remaining 47% unexplained.…”

Section: Discussionmentioning

confidence: 98%

“…Nevertheless, screening for PROS1 gene mutations has been performed in a number of studies. The proportion of cases where no mutations are detected varies widely between studies [10,17,23–33]. When all cases are pooled together, PROS1 gene mutations are not detected in 47% of PS deficient families.…”

Section: Introductionmentioning

confidence: 99%

Co‐segregation of the PROS1 locus and protein S deficiency in families having no detectable mutations in PROS1

Lanke

Johansson

Hillarp

et al. 2004

Journal of Thrombosis and Haemostasis

View full text Add to dashboard Cite

Summary. Inherited deficiency of protein S constitutes an important risk factor of venous thrombosis. Many reports have demonstrated that causative mutations in the protein S gene are found only in approximately 50% of the cases with protein S deficiency. It is uncertain whether the protein S gene is causative in all cases of protein S deficiency or if other genes are involved in cases where no mutation is identified. The aim of the current study was to determine whether haplotypes of the protein S gene cosegregate with the disease phenotype in cases where no mutations have been found. Eight protein S‐deficient families comprising 115 individuals where previous DNA sequencing had failed to detect any causative mutations were analyzed using four microsatellite markers in the protein S gene region. Co‐segregation between microsatellite haplotypes and protein S deficiency was found in seven of the investigated families, one family being uninformative. This suggests that the causative genetic defects are located in or close to the protein S gene in a majority of such cases where no mutations have been found.

show abstract

“…Type III deficiency is characterised by low levels of free PS and levels of total PS antigen within the normal range. Type I and type III phenotypes sometimes occur together within families (12), even though family members can carry the same underlying genetic defect (13)(14)(15)(16)(17).…”

Section: Introductionmentioning

confidence: 99%

Genetic and Phenotypic Variability between Families with Hereditary Protein S Deficiency

Rezende¹,

Lane²,

Zöller³

et al. 2002

Thromb Haemost

View full text Add to dashboard Cite

SummaryWhile many mutations thought to result in protein S (PS) deficiency are known, there have been few attempts to relate genotype expression with plasma phenotype. We have investigated the nature and consequence of PS gene (PROS1) mutations in 17 PS-deficient families who presented with mixed type I and type III phenotypes. Seven different mutations were found in nine families: delG-34 (STOP codon at –24), Val-24Glu, Arg49Cys, Asn217Ser, Gly295Val, +5 G to A intron j and His623Pro. PS wild type (PSWT) and the five missense mutants were transiently expressed in COS-1 cells. All mutants expressed lower (p<0.05) PS antigen compared to PSWT (100%). The mutants Val-24Glu, Gly295Val and His623Pro expressed very low/undetectable PS levels. The mutant Asn217Ser produced around 30% of PSWT, while the mutant Arg49Cys had the highest PS levels (around 50%). Metabolic labelling and pulse-chase experiments showed that all of the mutants had impaired secretion, but this was of variable severity. Also, enhanced intracellular degradation of unsecreted material was found for all mutants. There was a strong correspondence between plasma free PS levels in carriers of the mutations, secreted PS from transfected COS-1 cells and labelled PS from 24 h conditioned medium in pulse-chase experiment. We conclude that the magnitude of secretion defect depends on the nature of the PROS1 mutation and influences the level of free PS in plasma. It is likely that the severity of the secretion defect will determine the risk for venous thrombosis.

show abstract

Detection and characterization of seven novel protein S (PROS) gene lesions: evaluation of reverse transcript-polymerase chain reaction as a mutation screening strategy

Cited by 37 publications

References 29 publications

The prevalence of, and molecular defects underlying, inherited protein S deficiency in the general population

The prevalence of, and molecular defects underlying, inherited protein S deficiency in the general population

Co‐segregation of the PROS1 locus and protein S deficiency in families having no detectable mutations in PROS1

Genetic and Phenotypic Variability between Families with Hereditary Protein S Deficiency

Contact Info

Product

Resources

About