Comparative Genetic and Physiological Studies of the MAP Kinase Mpk1p from Kluyveromyces lactis and Saccharomyces cerevisiae

Kirchrath, Lutz; Lorberg, Anja; Schmitz, Hans‐Peter; Gengenbacher, Ute; Heinisch, Jürgen J.

doi:10.1006/jmbi.2000.3916

Cited by 58 publications

(70 citation statements)

References 51 publications

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“…If necessary, the isogenic diploid strain DHD5 (Kirchrath et al, 2000) was employed for gene deletion and gene fusion experiments and subsequently subjected to sporulation and tetrad analyses. Escherichia coli strain DH5αF (Gibco, BRL) was used as a bacterial host for plasmids, throughout.…”

Section: Strainsmentioning

confidence: 99%

“…As a basis to obtain lexA-RLM1-lacZ reporter constructs with different selectable markers, pHPS100 was used, a CEN-ARS vector carrying the TRP1 marker (Kirchrath et al, 2000). To substitute the TRP1 marker for HIS3, LEU2 or URA3, plasmids YDp-H, YDp-L and YDp-U …”

Section: Oligonucleotides Pcr Cassettes and Plasmidsmentioning

confidence: 99%

“…This then serves as a less laborious, albeit indirect, measure of pathway activity. We have previously placed all necessary components for this detection system on a single yeast CEN-ARS plasmid carrying a TRP1 selection marker (Kirchrath et al, 2000). However, a trp1 defect leads to drastically increased sensitivities of mutants defective in cell integrity signalling to a variety of stresses, such as addition of Calcofluor white or SDS (Queralt and Igual, 2005;unpub-lished results from our laboratory).…”

Section: Introductionmentioning

confidence: 99%

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The effect of tea tree oil and antifungal agents on a reporter for yeast cell integrity signalling

Straede

Corran

Bundy

et al. 2007

Yeast

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Cell integrity in Saccharomyces cerevisiae is ensured by a rigid cell wall whose synthesis is controlled by a highly conserved MAP kinase signal transduction cascade. Stress at the cell surface is detected by a set of sensors and ultimately transmitted through this cascade to the transcription factor Rlm1, which governs expression of many genes encoding enzymes of cell wall biosynthesis. We here report on a number of versatile reporter constructs which link activation of a hybrid, Rlm1-lexA, by the MAP kinase Mpk1/Slt2 to the expression of the bacterial lacZ gene. This system was adapted to automated microwell screening and shown to be activated by a number of compounds inhibiting cell wall biosynthesis or interfering with plasma membrane function. In addition, we tested tea tree oil and two of its purified constituents (α-terpineol, terpinen-4-ol) for their effects on growth and on cell integrity signalling using such reporter strains. Tea tree oil was found to inhibit growth of wild-type and slg1/wsc1 mutant cells at a threshold of approximately 0.1% v/v, with the purified compounds acting already at half these concentrations. A mid2 deletion displayed hyper-resistance. Tea tree oil also induces the signalling pathway in a dose-dependent manner.

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Section: Strainsmentioning

confidence: 99%

Section: Oligonucleotides Pcr Cassettes and Plasmidsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The effect of tea tree oil and antifungal agents on a reporter for yeast cell integrity signalling

Straede

Corran

Bundy

et al. 2007

Yeast

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“…The presence of transactivation domains in Slt2p might also reflect a role of this MAPK as a kinase co-activator. In support of this notion, Kirchrath et al, (2000) reported that a yeast strain expressing a C-terminal truncation of Slt2p (amino acids 1-322) as its only copy of this gene displayed sensitivity to the cell wall antagonist caffeine as well as diminished activation of an Rlm1p-based transcriptional reporter.…”

Section: Discussionmentioning

confidence: 89%

“…Interestingly, other fungal MAPKs involved in CWI signalling, such as Candida albicans Mkc1 (Navarro-García et al, 1998), Pichia pastoris Pim1p (Cosano et al, 2001) or Kluyveromyces lactis Mpk1p (Kirchrath et al, 2000), are able to induce transcriptional activation in one-hybrid assays, whereas the Erk1/2-like MAPKs Fus3p and Kss1p do not possess such transactivation ability (data not shown). The CWI MAPKs have in common a C-terminal non-catalytic extension, which is absent in the latter MAPKs.…”

Section: Discussionmentioning

confidence: 97%

Dissecting the transcriptional activation function of the cell wall integrity MAP kinase

Kim

Cosano

Levin

et al. 2007

Yeast

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The cell wall integrity signalling MAP kinase of Saccharomyces cerevisiae, Slt2p/ Mpk1p, is activated in response to cell wall stress. Slt2 and its mammalian orthologue ERK5 are unusual among MAP kinases, in that they possess the ability to activate transcription of a GAL1-lacZ reporter when fused to the DNA-binding domain of the Gal4 transcription factor. In this study, we demonstrate that transcriptional activation of a Gal4-Slt2p fusion is responsive to cell wall stress and requires phosphorylation of Slt2p. We identify two neighbouring but separable transcription activation domains within the C-terminal half of Slt2p. Additionally, we present data suggesting that intramolecular interactions controlled by phosphorylation of Slt2p regulate the function of these domains, which are masked by the N-terminal catalytic domain under inactive conditions. Finally, we demonstrate that Slt2p self-associates, probably through a glutamine-rich region within the C-terminal half of the protein.

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Comparative Genetic and Physiological Studies of the MAP Kinase Mpk1p from Kluyveromyces lactis and Saccharomyces cerevisiae

Cited by 58 publications

References 51 publications

The effect of tea tree oil and antifungal agents on a reporter for yeast cell integrity signalling

The effect of tea tree oil and antifungal agents on a reporter for yeast cell integrity signalling

Dissecting the transcriptional activation function of the cell wall integrity MAP kinase

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