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Clinical and functional properties of novel VHL mutation (X214L) consistent with Type 2A phenotype and low risk of renal cell carcinoma.

Sorrell AD, Lee S, Stolle C, Ellenhorn J, Grix A, Kaelin WG, Weitzel JN - Clin. Genet. (2011)

Bottom Line: In contrast, the variant was defective with respect to downregulation of JunB. pVHL X214L, like other pVHL variants associated with a low risk of clear cell renal carcinoma, largely preserves the ability to downregulate HIF.In contrast, this variant, like other pVHL variants linked to Type 2A disease, fails to suppress JunB.This underscores that JunB may play a role in the pathogenesis of Type 2A VHL disease.

View Article: PubMed Central - PubMed

Affiliation: City of Hope National Medical Center, Duarte, California 91010-3000, USA. asorrell@coh.org

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Immunoblot analysis using two VHL protein (pVHL)  renal carcinoma cell lines (786-O and RCC-4). Where indicated, cells were treated with 1mM of dimethyloxaloylglycine (DMOG), dissolved in dimethyl sulfoxide (DMSO), for 16 h to induce HIF2α protein. DMOG-induced HIF2α protein in the X214L mutant (X) and wt VHL (V) expressing cells. Compared to wt VHL, high-levels of JunB expression was seen in the X214L mutant expressing cell lines, pre- and post-DMOG exposure. No pre- and post-DMOG expression differences were detected in the cells expressing the empty vector (E). This data represents three independent experiments.
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fig03: Immunoblot analysis using two VHL protein (pVHL) renal carcinoma cell lines (786-O and RCC-4). Where indicated, cells were treated with 1mM of dimethyloxaloylglycine (DMOG), dissolved in dimethyl sulfoxide (DMSO), for 16 h to induce HIF2α protein. DMOG-induced HIF2α protein in the X214L mutant (X) and wt VHL (V) expressing cells. Compared to wt VHL, high-levels of JunB expression was seen in the X214L mutant expressing cell lines, pre- and post-DMOG exposure. No pre- and post-DMOG expression differences were detected in the cells expressing the empty vector (E). This data represents three independent experiments.

Mentions: Prior studies suggest that pVHL mutants linked to familial pheochromocytoma fail to downregulate the c-Jun antagonist JunB, attenuating apoptosis, and setting the stage for neoplastic transformation (10). Using two independent, pVHL , RCC lines (786-O and RCC-4), we found that the X214L pVHL mutant is associated with abnormal JunB expression, when compared to wild-type pVHL (Fig. 3). The electrophoretic mobility of the exogenous HA-tagged pVHL (X214L) was slower than that of HA-tagged wild-type pVHL, consistent with the presence of 14 additional amino acids. HIF2α induction patterns were assessed after treatment with dimethyloxaloylglycine (DMOG), a prolyl hydroxylase inhibitor, in cell lines expressing wild-type or X214L pVHL. The X214L mutant pVHL appeared to downregulate HIFα protein expression, in a canonical, hydroxylation-dependent, manner. In contrast, X214L was defective relative to wild-type pVHL with respect to downregulation of JunB. In sum, X214L mutant protein was associated with low HIF2α expression and high JunB expression, a pattern consistent with a low risk of kidney cancer and high risk of pheochromocytoma.


Clinical and functional properties of novel VHL mutation (X214L) consistent with Type 2A phenotype and low risk of renal cell carcinoma.

Sorrell AD, Lee S, Stolle C, Ellenhorn J, Grix A, Kaelin WG, Weitzel JN - Clin. Genet. (2011)

Immunoblot analysis using two VHL protein (pVHL)  renal carcinoma cell lines (786-O and RCC-4). Where indicated, cells were treated with 1mM of dimethyloxaloylglycine (DMOG), dissolved in dimethyl sulfoxide (DMSO), for 16 h to induce HIF2α protein. DMOG-induced HIF2α protein in the X214L mutant (X) and wt VHL (V) expressing cells. Compared to wt VHL, high-levels of JunB expression was seen in the X214L mutant expressing cell lines, pre- and post-DMOG exposure. No pre- and post-DMOG expression differences were detected in the cells expressing the empty vector (E). This data represents three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC2958253&req=5

fig03: Immunoblot analysis using two VHL protein (pVHL) renal carcinoma cell lines (786-O and RCC-4). Where indicated, cells were treated with 1mM of dimethyloxaloylglycine (DMOG), dissolved in dimethyl sulfoxide (DMSO), for 16 h to induce HIF2α protein. DMOG-induced HIF2α protein in the X214L mutant (X) and wt VHL (V) expressing cells. Compared to wt VHL, high-levels of JunB expression was seen in the X214L mutant expressing cell lines, pre- and post-DMOG exposure. No pre- and post-DMOG expression differences were detected in the cells expressing the empty vector (E). This data represents three independent experiments.
Mentions: Prior studies suggest that pVHL mutants linked to familial pheochromocytoma fail to downregulate the c-Jun antagonist JunB, attenuating apoptosis, and setting the stage for neoplastic transformation (10). Using two independent, pVHL , RCC lines (786-O and RCC-4), we found that the X214L pVHL mutant is associated with abnormal JunB expression, when compared to wild-type pVHL (Fig. 3). The electrophoretic mobility of the exogenous HA-tagged pVHL (X214L) was slower than that of HA-tagged wild-type pVHL, consistent with the presence of 14 additional amino acids. HIF2α induction patterns were assessed after treatment with dimethyloxaloylglycine (DMOG), a prolyl hydroxylase inhibitor, in cell lines expressing wild-type or X214L pVHL. The X214L mutant pVHL appeared to downregulate HIFα protein expression, in a canonical, hydroxylation-dependent, manner. In contrast, X214L was defective relative to wild-type pVHL with respect to downregulation of JunB. In sum, X214L mutant protein was associated with low HIF2α expression and high JunB expression, a pattern consistent with a low risk of kidney cancer and high risk of pheochromocytoma.

Bottom Line: In contrast, the variant was defective with respect to downregulation of JunB. pVHL X214L, like other pVHL variants associated with a low risk of clear cell renal carcinoma, largely preserves the ability to downregulate HIF.In contrast, this variant, like other pVHL variants linked to Type 2A disease, fails to suppress JunB.This underscores that JunB may play a role in the pathogenesis of Type 2A VHL disease.

View Article: PubMed Central - PubMed

Affiliation: City of Hope National Medical Center, Duarte, California 91010-3000, USA. asorrell@coh.org

Show MeSH
Related in: MedlinePlus