Modeling tumor predisposing FH mutations in yeast: Effects on fumarase activity, growth phenotype and gene expression profile

Antti Kokko, Sanna S.K. Ylisaukko-Oja, Maija Ht Kiuru, Maarit S. Takatalo, Paula Salmikangas, Jarno Tuimala, Diego Arango, Auli Karhu, Lauri A. Aaltonen, Jussi Jäntti

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Heterozygous mutations in the fumarase (FH) gene cause the tumor predisposition syndrome hereditary leiomyomatosis and renal cell cancer (MIM 605839). While most families segregate a benign phenotype of multiple leiomyomas, others display a phenotype with early-onset renal cancer and leiomyosarcoma. Modifier genes may play a role in this, but an alternative explanation is simple genotype-phenotype association. FH mutations predisposing to cancer appear to be truncating or in fully conserved amino acids, suggesting that mutations severely affecting FH activity might predispose to malignancy. In the present study, we analyzed 2 conserved fumarase mutations in yeast. H153R has been described in 3 cancer predisposition families; whereas all 3 reported K187R families have displayed the benign phenotype. Examining H153R and K187R should clarify whether cancer-related FH mutations differ from their benign phenotype-associated counterparts. Yeast strains containing the 2 mutations, and knockout and wild type (WT) references, were created and the growth phenotypes studied on selected carbon sources to assess mitochondrial function. Additionally, Fum1 protein production and activity were measured, and the strains were subjected to transcriptional profiling. On nonfermentable lactate medium, the fumarase knockout strains did not grow, whereas the mutants showed no differences, as compared to WT yeast. Although both mutant strains produced fumarase, a considerable decrease in enzyme activity was seen in mutants with respect to WT. Transcription of the majority of Krebs cycle enzymes was downregulated in response to mutations in fumarase. In conclusion, both mutants displayed some, albeit greatly reduced, fumarase activity. This activity was sufficient to support normal growth on nonfermentable carbon source, unlike the deletion phenotype, demonstrating the significance of the residual activity. The findings support the hypothesis that modifier gene(s), rather than phenotype-genotype effects, display a major role in determining tumor phenotypes in families segregating FH mutations.

Original languageEnglish (US)
Pages (from-to)1340-1345
Number of pages6
JournalInternational Journal of Cancer
Volume118
Issue number6
DOIs
StatePublished - Mar 15 2006
Externally publishedYes

Fingerprint

Fumarate Hydratase
Transcriptome
Yeasts
Phenotype
Mutation
Growth
Neoplasms
Modifier Genes
Carbon
Citric Acid Cycle
Leiomyosarcoma
Kidney Neoplasms
Genetic Association Studies
Leiomyoma
Enzymes
Lactic Acid
Down-Regulation
Genotype
Amino Acids

Keywords

  • Fumarase
  • HLRCC
  • Microarray
  • Yeast

ASJC Scopus subject areas

  • Medicine(all)
  • Oncology
  • Cancer Research

Cite this

Modeling tumor predisposing FH mutations in yeast : Effects on fumarase activity, growth phenotype and gene expression profile. / Kokko, Antti; Ylisaukko-Oja, Sanna S.K.; Kiuru, Maija Ht; Takatalo, Maarit S.; Salmikangas, Paula; Tuimala, Jarno; Arango, Diego; Karhu, Auli; Aaltonen, Lauri A.; Jäntti, Jussi.

In: International Journal of Cancer, Vol. 118, No. 6, 15.03.2006, p. 1340-1345.

Research output: Contribution to journalArticle

Kokko, A, Ylisaukko-Oja, SSK, Kiuru, MH, Takatalo, MS, Salmikangas, P, Tuimala, J, Arango, D, Karhu, A, Aaltonen, LA & Jäntti, J 2006, 'Modeling tumor predisposing FH mutations in yeast: Effects on fumarase activity, growth phenotype and gene expression profile', International Journal of Cancer, vol. 118, no. 6, pp. 1340-1345. https://doi.org/10.1002/ijc.21423
Kokko, Antti ; Ylisaukko-Oja, Sanna S.K. ; Kiuru, Maija Ht ; Takatalo, Maarit S. ; Salmikangas, Paula ; Tuimala, Jarno ; Arango, Diego ; Karhu, Auli ; Aaltonen, Lauri A. ; Jäntti, Jussi. / Modeling tumor predisposing FH mutations in yeast : Effects on fumarase activity, growth phenotype and gene expression profile. In: International Journal of Cancer. 2006 ; Vol. 118, No. 6. pp. 1340-1345.
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abstract = "Heterozygous mutations in the fumarase (FH) gene cause the tumor predisposition syndrome hereditary leiomyomatosis and renal cell cancer (MIM 605839). While most families segregate a benign phenotype of multiple leiomyomas, others display a phenotype with early-onset renal cancer and leiomyosarcoma. Modifier genes may play a role in this, but an alternative explanation is simple genotype-phenotype association. FH mutations predisposing to cancer appear to be truncating or in fully conserved amino acids, suggesting that mutations severely affecting FH activity might predispose to malignancy. In the present study, we analyzed 2 conserved fumarase mutations in yeast. H153R has been described in 3 cancer predisposition families; whereas all 3 reported K187R families have displayed the benign phenotype. Examining H153R and K187R should clarify whether cancer-related FH mutations differ from their benign phenotype-associated counterparts. Yeast strains containing the 2 mutations, and knockout and wild type (WT) references, were created and the growth phenotypes studied on selected carbon sources to assess mitochondrial function. Additionally, Fum1 protein production and activity were measured, and the strains were subjected to transcriptional profiling. On nonfermentable lactate medium, the fumarase knockout strains did not grow, whereas the mutants showed no differences, as compared to WT yeast. Although both mutant strains produced fumarase, a considerable decrease in enzyme activity was seen in mutants with respect to WT. Transcription of the majority of Krebs cycle enzymes was downregulated in response to mutations in fumarase. In conclusion, both mutants displayed some, albeit greatly reduced, fumarase activity. This activity was sufficient to support normal growth on nonfermentable carbon source, unlike the deletion phenotype, demonstrating the significance of the residual activity. The findings support the hypothesis that modifier gene(s), rather than phenotype-genotype effects, display a major role in determining tumor phenotypes in families segregating FH mutations.",
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