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Novel polymorphic AluYb8 insertion in the WNK1 gene is associated with blood pressure variation in Europeans.

Putku M, Kepp K, Org E, Sõber S, Comas D, Viigimaa M, Veldre G, Juhanson P, Hallast P, Tõnisson N, HYPertension in ESTonia (HYPEST)Shaw-Hawkins S, Caulfield MJ, BRItish Genetics of HyperTension (BRIGHT)Khusnutdinova E, Kožich V, Munroe PB, Laan M - Hum. Mutat. (2011)

Bottom Line: Meta-analysis across three European sample sets (n = 3,494; HYPEST, Estonians; BRIGHT, the British; CADCZ, Czech) detected significant association of the WNK1 AluYb8 insertion with blood pressure (BP; systolic BP, P = 4.03 × 10(-3) , effect 1.12; diastolic BP, P = 1.21 × 10(-2) , effect 0.67).Gender-stratified analysis revealed that this effect might be female-specific (n = 2,088; SBP, P = 1.99 × 10(-3) , effect 1.59; DBP P = 3.64 × 10(-4) , effect 1.23; resistant to Bonferroni correction), whereas no statistical support was identified for the association with male BP (n = 1,406).In leucocytes, the expressional proportions of the full-length WNK1 transcript and the splice-form skipping exon 11 were significantly shifted in AluYb8 carriers compared to noncarriers.

View Article: PubMed Central - PubMed

Affiliation: Human Molecular Genetics Research Group, Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia.

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Expression of (A) three WNK1 alternative splice-forms in blood leucocytes obtained from (B) heterozygous (Alu/−) and (C) homozygous (Alu/Alu) carriers of the WNK1 AluYb8 insertion in comparison with the wild-type homozygote without the insertion. A: Alternative splicing of WNK1 exons 10–13 is presented schematically according to Delaloy et al. [2003]. Black numbered boxes and horizontal lines represent exons and introns, respectively, and dotted lines indicate splicing events. B, C: Relative mRNA quantification of the targeted WNK1 splice-forms in leucocytes was performed with real-time RT-PCR (Taqman assay, HPRT as a reference gene). Relative expression of each targeted WNK1 splice-form in subjects with the AluYb8 insertion (Alu/Alu homozygotes, Alu/− heterozygotes) is shown using the quantity of the transcript in wild-type homozygotes (−/−) as a reference value (wt = 1). The presented relative expression levels represent the mean values of the three study subjects within the genotype group (each individual represented by six data points from replicate experiments). Bars represent standard error of the relative expression. P-Values reflecting the differences between groups were estimated by Wilcoxon rank sum test.
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fig02: Expression of (A) three WNK1 alternative splice-forms in blood leucocytes obtained from (B) heterozygous (Alu/−) and (C) homozygous (Alu/Alu) carriers of the WNK1 AluYb8 insertion in comparison with the wild-type homozygote without the insertion. A: Alternative splicing of WNK1 exons 10–13 is presented schematically according to Delaloy et al. [2003]. Black numbered boxes and horizontal lines represent exons and introns, respectively, and dotted lines indicate splicing events. B, C: Relative mRNA quantification of the targeted WNK1 splice-forms in leucocytes was performed with real-time RT-PCR (Taqman assay, HPRT as a reference gene). Relative expression of each targeted WNK1 splice-form in subjects with the AluYb8 insertion (Alu/Alu homozygotes, Alu/− heterozygotes) is shown using the quantity of the transcript in wild-type homozygotes (−/−) as a reference value (wt = 1). The presented relative expression levels represent the mean values of the three study subjects within the genotype group (each individual represented by six data points from replicate experiments). Bars represent standard error of the relative expression. P-Values reflecting the differences between groups were estimated by Wilcoxon rank sum test.

Mentions: Relative expression analysis of three WNK1 splice forms (ex+11+12, ex−11+12, and ex−11−12; Fig. 2A) was performed with real-time PCR. Primer-probe mix of the WNK1 transcript including exon 11 (ex+11+12; Hs01018312_m1, amplicon size 78 bp) and selected reference gene HPRT1 [Human HPRT1 (HGPRT) Endogenous Control (VIC/MGB Probe, Primer Limited, amplicon size 100 bp)] were purchased from Applied Biosystems, Inc. (Foster City, CA). Primers and probes for the WNK1 transcripts lacking exon 11 (ex−11+12) and both exons 11 and 12 (ex−11−12) were designed using Primer Express version 3.0 (Applied Biosystems Inc.). Oligonucleotide sequences are given in Supp. Table S4.


Novel polymorphic AluYb8 insertion in the WNK1 gene is associated with blood pressure variation in Europeans.

Putku M, Kepp K, Org E, Sõber S, Comas D, Viigimaa M, Veldre G, Juhanson P, Hallast P, Tõnisson N, HYPertension in ESTonia (HYPEST)Shaw-Hawkins S, Caulfield MJ, BRItish Genetics of HyperTension (BRIGHT)Khusnutdinova E, Kožich V, Munroe PB, Laan M - Hum. Mutat. (2011)

Expression of (A) three WNK1 alternative splice-forms in blood leucocytes obtained from (B) heterozygous (Alu/−) and (C) homozygous (Alu/Alu) carriers of the WNK1 AluYb8 insertion in comparison with the wild-type homozygote without the insertion. A: Alternative splicing of WNK1 exons 10–13 is presented schematically according to Delaloy et al. [2003]. Black numbered boxes and horizontal lines represent exons and introns, respectively, and dotted lines indicate splicing events. B, C: Relative mRNA quantification of the targeted WNK1 splice-forms in leucocytes was performed with real-time RT-PCR (Taqman assay, HPRT as a reference gene). Relative expression of each targeted WNK1 splice-form in subjects with the AluYb8 insertion (Alu/Alu homozygotes, Alu/− heterozygotes) is shown using the quantity of the transcript in wild-type homozygotes (−/−) as a reference value (wt = 1). The presented relative expression levels represent the mean values of the three study subjects within the genotype group (each individual represented by six data points from replicate experiments). Bars represent standard error of the relative expression. P-Values reflecting the differences between groups were estimated by Wilcoxon rank sum test.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3298642&req=5

fig02: Expression of (A) three WNK1 alternative splice-forms in blood leucocytes obtained from (B) heterozygous (Alu/−) and (C) homozygous (Alu/Alu) carriers of the WNK1 AluYb8 insertion in comparison with the wild-type homozygote without the insertion. A: Alternative splicing of WNK1 exons 10–13 is presented schematically according to Delaloy et al. [2003]. Black numbered boxes and horizontal lines represent exons and introns, respectively, and dotted lines indicate splicing events. B, C: Relative mRNA quantification of the targeted WNK1 splice-forms in leucocytes was performed with real-time RT-PCR (Taqman assay, HPRT as a reference gene). Relative expression of each targeted WNK1 splice-form in subjects with the AluYb8 insertion (Alu/Alu homozygotes, Alu/− heterozygotes) is shown using the quantity of the transcript in wild-type homozygotes (−/−) as a reference value (wt = 1). The presented relative expression levels represent the mean values of the three study subjects within the genotype group (each individual represented by six data points from replicate experiments). Bars represent standard error of the relative expression. P-Values reflecting the differences between groups were estimated by Wilcoxon rank sum test.
Mentions: Relative expression analysis of three WNK1 splice forms (ex+11+12, ex−11+12, and ex−11−12; Fig. 2A) was performed with real-time PCR. Primer-probe mix of the WNK1 transcript including exon 11 (ex+11+12; Hs01018312_m1, amplicon size 78 bp) and selected reference gene HPRT1 [Human HPRT1 (HGPRT) Endogenous Control (VIC/MGB Probe, Primer Limited, amplicon size 100 bp)] were purchased from Applied Biosystems, Inc. (Foster City, CA). Primers and probes for the WNK1 transcripts lacking exon 11 (ex−11+12) and both exons 11 and 12 (ex−11−12) were designed using Primer Express version 3.0 (Applied Biosystems Inc.). Oligonucleotide sequences are given in Supp. Table S4.

Bottom Line: Meta-analysis across three European sample sets (n = 3,494; HYPEST, Estonians; BRIGHT, the British; CADCZ, Czech) detected significant association of the WNK1 AluYb8 insertion with blood pressure (BP; systolic BP, P = 4.03 × 10(-3) , effect 1.12; diastolic BP, P = 1.21 × 10(-2) , effect 0.67).Gender-stratified analysis revealed that this effect might be female-specific (n = 2,088; SBP, P = 1.99 × 10(-3) , effect 1.59; DBP P = 3.64 × 10(-4) , effect 1.23; resistant to Bonferroni correction), whereas no statistical support was identified for the association with male BP (n = 1,406).In leucocytes, the expressional proportions of the full-length WNK1 transcript and the splice-form skipping exon 11 were significantly shifted in AluYb8 carriers compared to noncarriers.

View Article: PubMed Central - PubMed

Affiliation: Human Molecular Genetics Research Group, Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia.

Show MeSH
Related in: MedlinePlus