A genetic interaction between NDPK and AMPK in Dictyostelium discoideum that affects motility, growth and development

Sarah J. Annesley, Ruzica Bago, Anil Mehta, Paul R. Fisher

    Research output: Contribution to journalArticle

    10 Citations (Scopus)

    Abstract

    Many of the expanding roles of nucleoside diphosphate kinase have been attributed to its ability to interact with other proteins. One proposal is an interaction with the cellular energy sensor AMP-activated protein kinase, and here, we apply the simple eukaryotic organism, Dictyostelium discoideum as a test model. Stable cotransformants were created in which NDPK expression was knocked down by antisense inhibition, and AMPK activity was chronically elevated either by constitutive overexpression of its active, catalytic domain (AMPK alpha T) or as a result of mitochondrial dysfunction (created by antisense inhibition of expression of a mitochondrial chaperone protein, chaperonin 60). To investigate a biochemical interaction, transformants were created which contained constructs expressing FLAG-NDPK and hexahistidine-tagged full-length AMPK or AMPK alpha T. The protein extract from these transformants was used in coimmunoprecipitations. Knock down of NDPK expression suppressed the phenotypic defects that are caused by AMPK hyperactivity resulting either from overexpression of AMPK alpha T or from mitochondrial dysfunction. These included rescue of defects in slug phototaxis, fruiting body morphology and growth in a liquid medium. Coimmunoprecipitation experiments failed to demonstrate a biochemical interaction between the two proteins. The results demonstrate a genetic interaction between NDPK and AMPK in Dictyostelium in that NDPK is required for the phenotypic effects of activated AMPK. Coimmunoprecipitations suggest that this interaction is not mediated by a direct interaction between the two proteins.

    Original languageEnglish
    Pages (from-to)341-349
    Number of pages9
    JournalNaunyn-Schmiedeberg's Archives of Pharmacology
    Volume384
    Issue number4-5
    DOIs
    Publication statusPublished - Oct 2011

    Keywords

    • NDPK
    • nm23
    • AMPK
    • Dictyostelium
    • NUCLEOSIDE DIPHOSPHATE KINASE
    • X-RAY-STRUCTURE
    • SLUG PHOTOTAXIS
    • CELL-ADHESION
    • PHOSPHORYLATION
    • DROSOPHILA
    • EPITHELIA
    • MUTATIONS
    • BEHAVIOR
    • NM23-H1

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