Mutations at the hydrophobic core affect Hal3 trimer stability, reducing its Ppz1 inhibitory capacity but not its PPCDC moonlighting function

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dc.contributor.authorSantolaria, Carlosen_ZA
dc.contributor.authorVelazquez, Diegoen_ZA
dc.contributor.authorStrauss, Ericken_ZA
dc.contributor.authorArino, Joaquinen_ZA
dc.date.accessioned2020-03-17T07:21:00Z
dc.date.available2020-03-17T07:21:00Z
dc.date.issued2018-10-02en_ZA
dc.descriptionCITATION: Santolaria, C., et al. 2018. Mutations at the hydrophobic core affect Hal3 trimer stability, reducing its Ppz1 inhibitory capacity but not its PPCDC moonlighting function. Scientific Reports, 8:14701, doi:10.1038/s41598-018-32979-x.
dc.descriptionThe original publication is available at https://www.nature.com
dc.description.abstractS. cerevisiae Hal3 (ScHal3) is a moonlighting protein that, is in its monomeric state, regulates the Ser/Thr protein phosphatase Ppz1, but also joins ScCab3 (and in some instances the Hal3 paralog Vhs3) to form an unusual heterotrimeric phosphopantothenoylcysteine decarboxylase (PPCDC) enzyme. PPCDC is required for CoA biosynthesis and in most eukaryotes is a homotrimeric complex with three identical catalytic sites at the trimer interfaces. However, in S. cerevisiae the heterotrimeric arrangement results in a single functional catalytic center. Importantly, the specific structural determinants that direct Hal3’s oligomeric state and those required for Ppz1 inhibition remain largely unknown. We mutagenized residues in the predicted hydrophobic core of ScHal3 (L403–L405) and the plant Arabidopsis thaliana Hal3 (AtHal3, G115–L117) oligomers and characterized their properties as PPCDC components and, for ScHal3, also as Ppz1 inhibitor. We found that in AtHal3 these changes do not affect trimerization or PPCDC function. Similarly, mutation of ScHal3 L403 has no effect. In contrast, ScHal3 L405E fails to form homotrimers, but retains the capacity to bind Cab3—explaining its ability to rescue a hal3 vhs3 synthetically lethal mutation. Remarkably, the L405E mutation decreases Hal3’s ability to interact with and to inhibit Ppz1, confirming the importance of the oligomer/monomer equilibrium in Hal3’s Ppz1 regulating function.en_ZA
dc.description.versionPublisher's version
dc.format.extent12 pages : illustrationsen_ZA
dc.identifier.citationSantolaria, C., et al. 2018. Mutations at the hydrophobic core affect Hal3 trimer stability, reducing its Ppz1 inhibitory capacity but not its PPCDC moonlighting function. Scientific Reports, 8:14701, doi:10.1038/s41598-018-32979-x
dc.identifier.issn2045-2322 (online)
dc.identifier.otherdoi:10.1038/s41598-018-32979-x
dc.identifier.urihttp://hdl.handle.net/10019.1/107628
dc.language.isoen_ZAen_ZA
dc.publisherNature Research
dc.rights.holderAuthors retain copyright
dc.subjectPhosphoprotein phosphatasesen_ZA
dc.subjectSaccharomyces cerevisiae --- Biotechnologyen_ZA
dc.subjectProteins --- Analysisen_ZA
dc.titleMutations at the hydrophobic core affect Hal3 trimer stability, reducing its Ppz1 inhibitory capacity but not its PPCDC moonlighting functionen_ZA
dc.typeArticleen_ZA
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