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 Kiuru, Maarit S. Takatalo, Paula Salmikangas, Jarno Tuimala, Diego Arango, Auli Karhu, Lauri A. Aaltonen, Jussi Jäntti (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

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
Pages (from-to)1340-1345
Number of pages6
JournalInternational Journal of Cancer
Volume118
Issue number6
DOIs
Publication statusPublished - 2006
MoE publication typeA1 Journal article-refereed

Fingerprint

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

Keywords

  • HLRCC
  • fumarase
  • yeasts
  • microarray

Cite this

Kokko, Antti ; Ylisaukko-oja, Sanna S.K. ; Kiuru, Maija ; 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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title = "Modeling tumor predisposing FH mutations in yeast: Effects on fumarase activity, growth phenotype and gene expression profile",
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.",
keywords = "HLRCC, fumarase, yeasts, microarray",
author = "Antti Kokko and Ylisaukko-oja, {Sanna S.K.} and Maija Kiuru and Takatalo, {Maarit S.} and Paula Salmikangas and Jarno Tuimala and Diego Arango and Auli Karhu and Aaltonen, {Lauri A.} and Jussi J{\"a}ntti",
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Kokko, A, Ylisaukko-oja, SSK, Kiuru, M, 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

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; Takatalo, Maarit S.; Salmikangas, Paula; Tuimala, Jarno; Arango, Diego; Karhu, Auli; Aaltonen, Lauri A.; Jäntti, Jussi (Corresponding Author).

In: International Journal of Cancer, Vol. 118, No. 6, 2006, p. 1340-1345.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Modeling tumor predisposing FH mutations in yeast

T2 - Effects on fumarase activity, growth phenotype and gene expression profile

AU - Kokko, Antti

AU - Ylisaukko-oja, Sanna S.K.

AU - Kiuru, Maija

AU - Takatalo, Maarit S.

AU - Salmikangas, Paula

AU - Tuimala, Jarno

AU - Arango, Diego

AU - Karhu, Auli

AU - Aaltonen, Lauri A.

AU - Jäntti, Jussi

PY - 2006

Y1 - 2006

N2 - 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.

AB - 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.

KW - HLRCC

KW - fumarase

KW - yeasts

KW - microarray

U2 - 10.1002/ijc.21423

DO - 10.1002/ijc.21423

M3 - Article

VL - 118

SP - 1340

EP - 1345

JO - International Journal of Cancer

JF - International Journal of Cancer

SN - 0020-7136

IS - 6

ER -