Elongator function depends on antagonistic regulation by casein kinase Hrr25 and protein phosphatase Sit4

Constance Mehlgarten, Daniel Jablonowski, Karin D. Breunig, Michael J. R. Stark, Raffael Schaffrath

    Research output: Contribution to journalArticle

    36 Citations (Scopus)

    Abstract

    P>In yeast, the role for the Elongator complex in tRNA anticodon modification is affected by phosphorylation of Elongator subunit Elp1. Thus, hyperphosphorylation of Elp1 due to inactivation of protein phosphatase Sit4 correlates with Elongator-minus phenotypes including resistance towards zymocin, a tRNase cleaving anticodons of Elongator-dependent tRNAs. Here we show that zymocin resistance of casein kinase hrr25 mutants associates with hypophosphorylation of Elp1 and that nonsense suppression by the Elongator-dependent SUP4 tRNA is abolished in hrr25 or sit4 mutants. Thus changes that perturb the evenly balanced ratio between hyper- and hypophosphorylated Elp1 forms present in wild-type cells lead to Elongator inactivation. Antagonistic roles for Hrr25 and Sit4 in Elongator function are further supported by our data that Sit4 inactivation is capable of restoring both zymocin sensitivity and normal ratios between the two Elp1 forms in hrr25 mutants. Hrr25 binds to Elongator in a fashion dependent on Elongator partner Kti12. Like sit4 mutants, overexpression of Kti12 triggers Elp1 hyperphosphorylation. Intriguingly, this effect of Kti12 is blocked by hrr25 mutations, which also show enhanced binding of Kti12 to Elongator. Collectively, our data suggest that rather than directly targeting Elp1, the Hrr25 kinase indirectly affects Elp1 phosphorylation states through control of Sit4-dependent dephosphorylation of Elp1.

    Original languageEnglish
    Pages (from-to)869-881
    Number of pages13
    JournalMolecular Microbiology
    Volume73
    Issue number5
    DOIs
    Publication statusPublished - Sep 2009

    Keywords

    • KLUYVEROMYCES-LACTIS ZYMOCIN
    • RNA-POLYMERASE-II
    • SACCHAROMYCES-CEREVISIAE
    • FAMILIAL DYSAUTONOMIA
    • YEAST PROTEOME
    • BUDDING YEAST
    • ORGAN GROWTH
    • GAMMA-TOXIN
    • COMPLEX
    • GENES

    Cite this

    Mehlgarten, Constance ; Jablonowski, Daniel ; Breunig, Karin D. ; Stark, Michael J. R. ; Schaffrath, Raffael. / Elongator function depends on antagonistic regulation by casein kinase Hrr25 and protein phosphatase Sit4. In: Molecular Microbiology. 2009 ; Vol. 73, No. 5. pp. 869-881.
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    title = "Elongator function depends on antagonistic regulation by casein kinase Hrr25 and protein phosphatase Sit4",
    abstract = "P>In yeast, the role for the Elongator complex in tRNA anticodon modification is affected by phosphorylation of Elongator subunit Elp1. Thus, hyperphosphorylation of Elp1 due to inactivation of protein phosphatase Sit4 correlates with Elongator-minus phenotypes including resistance towards zymocin, a tRNase cleaving anticodons of Elongator-dependent tRNAs. Here we show that zymocin resistance of casein kinase hrr25 mutants associates with hypophosphorylation of Elp1 and that nonsense suppression by the Elongator-dependent SUP4 tRNA is abolished in hrr25 or sit4 mutants. Thus changes that perturb the evenly balanced ratio between hyper- and hypophosphorylated Elp1 forms present in wild-type cells lead to Elongator inactivation. Antagonistic roles for Hrr25 and Sit4 in Elongator function are further supported by our data that Sit4 inactivation is capable of restoring both zymocin sensitivity and normal ratios between the two Elp1 forms in hrr25 mutants. Hrr25 binds to Elongator in a fashion dependent on Elongator partner Kti12. Like sit4 mutants, overexpression of Kti12 triggers Elp1 hyperphosphorylation. Intriguingly, this effect of Kti12 is blocked by hrr25 mutations, which also show enhanced binding of Kti12 to Elongator. Collectively, our data suggest that rather than directly targeting Elp1, the Hrr25 kinase indirectly affects Elp1 phosphorylation states through control of Sit4-dependent dephosphorylation of Elp1.",
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    author = "Constance Mehlgarten and Daniel Jablonowski and Breunig, {Karin D.} and Stark, {Michael J. R.} and Raffael Schaffrath",
    year = "2009",
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    language = "English",
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    Elongator function depends on antagonistic regulation by casein kinase Hrr25 and protein phosphatase Sit4. / Mehlgarten, Constance; Jablonowski, Daniel; Breunig, Karin D.; Stark, Michael J. R.; Schaffrath, Raffael.

    In: Molecular Microbiology, Vol. 73, No. 5, 09.2009, p. 869-881.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Elongator function depends on antagonistic regulation by casein kinase Hrr25 and protein phosphatase Sit4

    AU - Mehlgarten, Constance

    AU - Jablonowski, Daniel

    AU - Breunig, Karin D.

    AU - Stark, Michael J. R.

    AU - Schaffrath, Raffael

    PY - 2009/9

    Y1 - 2009/9

    N2 - P>In yeast, the role for the Elongator complex in tRNA anticodon modification is affected by phosphorylation of Elongator subunit Elp1. Thus, hyperphosphorylation of Elp1 due to inactivation of protein phosphatase Sit4 correlates with Elongator-minus phenotypes including resistance towards zymocin, a tRNase cleaving anticodons of Elongator-dependent tRNAs. Here we show that zymocin resistance of casein kinase hrr25 mutants associates with hypophosphorylation of Elp1 and that nonsense suppression by the Elongator-dependent SUP4 tRNA is abolished in hrr25 or sit4 mutants. Thus changes that perturb the evenly balanced ratio between hyper- and hypophosphorylated Elp1 forms present in wild-type cells lead to Elongator inactivation. Antagonistic roles for Hrr25 and Sit4 in Elongator function are further supported by our data that Sit4 inactivation is capable of restoring both zymocin sensitivity and normal ratios between the two Elp1 forms in hrr25 mutants. Hrr25 binds to Elongator in a fashion dependent on Elongator partner Kti12. Like sit4 mutants, overexpression of Kti12 triggers Elp1 hyperphosphorylation. Intriguingly, this effect of Kti12 is blocked by hrr25 mutations, which also show enhanced binding of Kti12 to Elongator. Collectively, our data suggest that rather than directly targeting Elp1, the Hrr25 kinase indirectly affects Elp1 phosphorylation states through control of Sit4-dependent dephosphorylation of Elp1.

    AB - P>In yeast, the role for the Elongator complex in tRNA anticodon modification is affected by phosphorylation of Elongator subunit Elp1. Thus, hyperphosphorylation of Elp1 due to inactivation of protein phosphatase Sit4 correlates with Elongator-minus phenotypes including resistance towards zymocin, a tRNase cleaving anticodons of Elongator-dependent tRNAs. Here we show that zymocin resistance of casein kinase hrr25 mutants associates with hypophosphorylation of Elp1 and that nonsense suppression by the Elongator-dependent SUP4 tRNA is abolished in hrr25 or sit4 mutants. Thus changes that perturb the evenly balanced ratio between hyper- and hypophosphorylated Elp1 forms present in wild-type cells lead to Elongator inactivation. Antagonistic roles for Hrr25 and Sit4 in Elongator function are further supported by our data that Sit4 inactivation is capable of restoring both zymocin sensitivity and normal ratios between the two Elp1 forms in hrr25 mutants. Hrr25 binds to Elongator in a fashion dependent on Elongator partner Kti12. Like sit4 mutants, overexpression of Kti12 triggers Elp1 hyperphosphorylation. Intriguingly, this effect of Kti12 is blocked by hrr25 mutations, which also show enhanced binding of Kti12 to Elongator. Collectively, our data suggest that rather than directly targeting Elp1, the Hrr25 kinase indirectly affects Elp1 phosphorylation states through control of Sit4-dependent dephosphorylation of Elp1.

    KW - KLUYVEROMYCES-LACTIS ZYMOCIN

    KW - RNA-POLYMERASE-II

    KW - SACCHAROMYCES-CEREVISIAE

    KW - FAMILIAL DYSAUTONOMIA

    KW - YEAST PROTEOME

    KW - BUDDING YEAST

    KW - ORGAN GROWTH

    KW - GAMMA-TOXIN

    KW - COMPLEX

    KW - GENES

    U2 - 10.1111/j.1365-2958.2009.06811.x

    DO - 10.1111/j.1365-2958.2009.06811.x

    M3 - Article

    VL - 73

    SP - 869

    EP - 881

    JO - Molecular Microbiology

    JF - Molecular Microbiology

    SN - 0950-382X

    IS - 5

    ER -