Modeling tumor predisposing FH mutations in yeast

effects on fumarase activity, growth phenotype and gene expression profile

Antti Kokko, Sanna Ylisaukko-oja, Maija Kiuru, Maarit Takatalo, Paula Salmikangas, Jarno Tuimala, Diego Arango, Auli Karhu, Lauri A Aaltonen, Jussi Jäntti

    Research output: Contribution to journalArticleScientificpeer-review

    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. (c) 2005 Wiley-Liss, Inc."
    Original languageEnglish
    JournalInternational Journal of Cancer
    Volume118
    Issue number6
    Pages (from-to)1340-1345
    Number of pages6
    ISSN0020-7136
    DOIs
    Publication statusPublished - 2006
    MoE publication typeA1 Journal article-refereed

    Fields of Science

    • 311 Basic medicine

    Cite this

    Kokko, A., Ylisaukko-oja, S., Kiuru, M., Takatalo, M., Salmikangas, P., Tuimala, J., ... Jäntti, J. (2006). Modeling tumor predisposing FH mutations in yeast: effects on fumarase activity, growth phenotype and gene expression profile. International Journal of Cancer, 118(6), 1340-1345. https://doi.org/10.1002/ijc.21423
    Kokko, Antti ; Ylisaukko-oja, Sanna ; Kiuru, Maija ; Takatalo, Maarit ; 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.
    @article{036c205a88654647b26bd14a67b12556,
    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. (c) 2005 Wiley-Liss, Inc.{"}",
    keywords = "311 Basic medicine",
    author = "Antti Kokko and Sanna Ylisaukko-oja and Maija Kiuru and Maarit Takatalo and Paula Salmikangas and Jarno Tuimala and Diego Arango and Auli Karhu and Aaltonen, {Lauri A} and Jussi J{\"a}ntti",
    year = "2006",
    doi = "10.1002/ijc.21423",
    language = "English",
    volume = "118",
    pages = "1340--1345",
    journal = "International Journal of Cancer",
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    Kokko, A, Ylisaukko-oja, S, Kiuru, M, Takatalo, M, 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; Kiuru, Maija; Takatalo, Maarit; Salmikangas, Paula; Tuimala, Jarno; Arango, Diego; Karhu, Auli; Aaltonen, Lauri A; Jäntti, Jussi.

    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

    AU - Kiuru, Maija

    AU - Takatalo, Maarit

    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. (c) 2005 Wiley-Liss, Inc."

    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. (c) 2005 Wiley-Liss, Inc."

    KW - 311 Basic medicine

    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 -