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Saccharomyces cerevisiae YCRO17c/CWH43 encodes a putative sensor/transporter protein upstream of the BCK2 branch of the PKC1-dependent cell wall integrity pathway

Abstract : TheSaccharomyces cerevisiae cwh43-2mutant, originally isolated for its Calcofluor whitehypersensitivity, displays several cell wall defects similar to mutants in the PKC1–MPK1pathway, including a growth defect and increased release ofb-1,6-glucan andb-glucosylated proteins into the growth medium at increased temperatures. The cloning ofCWH43showed that it corresponds toYCR017c and encodes a protein with 14–16transmembrane segments containing several putative phosphorylation and glycosylationsites. The N-terminal part of the amino acid sequence of Cwh43p shows 40% similaritywith the mammalian FRAG1, a membrane protein that activates the fibroblast growthfactor receptor of rat osteosarcoma (FGFR2-ROS) and with protein sequences of fouruncharacterized ORFs fromCaenorhabditis elegansand one fromDrosophila melanoga-ster. The C-terminus of Cwh43p shows low similarities with a xylose permease ofBacillusmegateriumand with putative sugar transporter fromD. melanogaster, and has 52%similarity with a protein sequence from aSchizosaccharomyces pombecDNA. ACwh43–GFP fusion protein suggested a plasma membrane localization, althoughlocalization to the internal structure of the cells could not be excluded, and it concentratesto the bud tip of small budded cells and to the neck of dividing cells. Deletion ofCWH43resulted in cell wall defects less pronounced than those of thecwh43-2mutant. This allele-specific phenotype appears to be due to a G–R substitution at position 57 in a highlyconserved region of the protein. Genetic analysis placesCWH43upstream of theBCK2branch of the PKC1 signalling pathway, sincecwh43mutations were synthetic lethal withpkc1deletion, whereas thecwh43defects could be rescued by overexpression ofBCK2andnot by high-copy-number expression of genes encoding downstream proteins of thePKC1pathway.However, unlikeBCK2, whose disruption in acln3mutant resulted in growtharrest in G1, no growth defect was observed in a doublecwh43 cln3mutants. Takentogether, it is proposed thatCWH43encodes a protein with putative sensor andtransporter domains acting in parallel to the main PKC1-dependent cell wall integritypathway, and that this gene has evolved into two distinct genes in higher eukaryotes.
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https://hal.inrae.fr/hal-02937544
Déposant : Hélène Martin-Yken <>
Soumis le : lundi 14 septembre 2020 - 10:26:50
Dernière modification le : mercredi 16 septembre 2020 - 09:45:48

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Hélène Martin-Yken, Adilia Dagkessamanskaia, Piet de Groot, Arthur Ram, Frans Klis, et al.. Saccharomyces cerevisiae YCRO17c/CWH43 encodes a putative sensor/transporter protein upstream of the BCK2 branch of the PKC1-dependent cell wall integrity pathway. Yeast, Wiley, 2001, 18 (9), pp.827-840. ⟨10.1002/yea.731⟩. ⟨hal-02937544⟩

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