Tolerant industrial yeast Saccharomyces cerevisiae posses a more robust cell wall integrity signaling pathway against 2-furaldehyde and 5-(hydroxymethyl)-2-furaldehyde

ABSTRACT Pumilio family (PUF) proteins affect specific genes by binding to, and inhibiting the translation or stability of, their transcripts. The PUF domain is required and sufficient for this function. One Saccharomyces cerevisiae PUF protein, Mpt5p (also called Puf5p or Uth4p), promotes stress tolerance and replicative life span (the maximum number of doublings a mother cell can undergo before entering into senescence) by an unknown mechanism thought to partly overlap with, but to be independent of, the cell wall integrity (CWI) pathway. Here, we found that mpt5Δ mutants also display a short chronological life span (the time cells stay alive in saturated cultures in synthetic medium), a defect that is suppressed by activation of CWI signaling. We found that Mpt5p is an upstream activator of the CWI pathway: mpt5Δ mutants display the appropriate phenotypes and genetic interactions, display low basal activity of the pathway, and are defective in activation of the pathway upon thermal stress. A set of mRNAs that specifically bind to Mpt5p was recently reported. One such putative target, LRG1, encodes a GTPase-activating protein for Rho1p that directly links Mpt5p to CWI signaling: Lrg1p inhibits CWI signaling, LRG1 mRNA contains a consensus Mpt5p-binding site in its putative 3′ untranslated region, loss of Lrg1p suppresses the temperature sensitivity and CWI signaling defects of mpt5Δ mutants, and LRG1 mRNA abundance is inhibited by Mpt5p. We conclude that Mpt5p is required for normal replicative and chronological life spans and that the CWI pathway is a key and direct downstream target of this PUF protein.

Download Full-text

Genetic dissection of the signaling pathway required for the cell wall integrity checkpoint

Journal of Cell Science ◽

10.1242/jcs.219063 ◽

2018 ◽

Vol 131 (13) ◽

pp. jcs219063 ◽

Cited By ~ 2

Author(s):

Yuko Sukegawa ◽

Takahiro Negishi ◽

Yo Kikuchi ◽

Keiko Ishii ◽

Miyuki Imanari ◽

...

Keyword(s):

Cell Wall ◽

Signaling Pathway ◽

Cell Wall Integrity ◽

Genetic Dissection

Download Full-text

Pathway-based signature transcriptional profiles as tolerance phenotypes for the adapted industrial yeast Saccharomyces cerevisiae resistant to furfural and HMF

Applied Microbiology and Biotechnology ◽

10.1007/s00253-020-10434-0 ◽

2020 ◽

Vol 104 (8) ◽

pp. 3473-3492 ◽

Cited By ~ 4

Author(s):

Z. Lewis Liu ◽

Menggen Ma

Keyword(s):

Saccharomyces Cerevisiae ◽

Industrial Yeast ◽

Yeast Saccharomyces Cerevisiae ◽

Transcriptional Profiles

Download Full-text

Cell fusion in yeast is negatively regulated by components of the cell wall integrity pathway

Molecular Biology of the Cell ◽

10.1091/mbc.e18-04-0236 ◽

2019 ◽

Vol 30 (4) ◽

pp. 441-452 ◽

Cited By ~ 2

Author(s):

Allison E. Hall ◽

Mark D. Rose

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Death ◽

Cell Wall ◽

Yeast Cell ◽

Cell Fusion ◽

Cell Walls ◽

Cell Wall Integrity ◽

Fusion Genes ◽

Cell Wall Degradation ◽

Cell Wall Integrity Pathway

During mating, Saccharomyces cerevisiae cells must degrade the intervening cell wall to allow fusion of the partners. Because improper timing or location of cell wall degradation would cause lysis, the initiation of cell fusion must be highly regulated. Here, we find that yeast cell fusion is negatively regulated by components of the cell wall integrity (CWI) pathway. Loss of the cell wall sensor, MID2, specifically causes “mating-induced death” after pheromone exposure. Mating-induced death is suppressed by mutations in cell fusion genes ( FUS1, FUS2, RVS161, CDC42), implying that mid2Δ cells die from premature fusion without a partner. Consistent with premature fusion, mid2Δ shmoos had thinner cell walls and lysed at the shmoo tip. Normally, Cdc42p colocalizes with Fus2p to form a focus only when mating cells are in contact (prezygotes) and colocalization is required for cell fusion. However, Cdc42p was aberrantly colocalized with Fus2p to form a focus in mid2Δ shmoos. A hyperactive allele of the CWI kinase Pkc1p ( PKC1*) caused decreased cell fusion and Cdc42p localization in prezygotes. In shmoos, PKC1* increased Cdc42p localization; however, it was not colocalized with Fus2p or associated with cell death. We conclude that Mid2p and Pkc1p negatively regulate cell fusion via Cdc42p and Fus2p.

Download Full-text

Purification of profilin from Saccharomyces cerevisiae and analysis of profilin-deficient cells.

The Journal of Cell Biology ◽

10.1083/jcb.110.1.105 ◽

1990 ◽

Vol 110 (1) ◽

pp. 105-114 ◽

Cited By ~ 153

Author(s):

B K Haarer ◽

S H Lillie ◽

A E Adams ◽

V Magdolen ◽

W Bandlow ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Wall ◽

Actin Polymerization ◽

Yeast Cells ◽

Predicted Amino Acid Sequence ◽

Temperature Sensitive ◽

Wild Type ◽

Yeast Saccharomyces Cerevisiae ◽

Ellipsoidal Shape ◽

Hydrolysis Of

We have isolated profilin from yeast (Saccharomyces cerevisiae) and have microsequenced a portion of the protein to confirm its identity; the region microsequenced agrees with the predicted amino acid sequence from a profilin gene recently isolated from S. cerevisiae (Magdolen, V., U. Oechsner, G. Müller, and W. Bandlow. 1988. Mol. Cell. Biol. 8:5108-5115). Yeast profilin resembles profilins from other organisms in molecular mass and in the ability to bind to polyproline, retard the rate of actin polymerization, and inhibit hydrolysis of ATP by monomeric actin. Using strains that carry disruptions or deletions of the profilin gene, we have found that, under appropriate conditions, cells can survive without detectable profilin. Such cells grow slowly, are temperature sensitive, lose the normal ellipsoidal shape of yeast cells, often become multinucleate, and generally grow much larger than wild-type cells. In addition, these cells exhibit delocalized deposition of cell wall chitin and have dramatically altered actin distributions.

Download Full-text

Cross Talk between the Cell Wall Integrity and Cyclic AMP/Protein Kinase A Pathways in Cryptococcus neoformans

mBio ◽

10.1128/mbio.01573-14 ◽

2014 ◽

Vol 5 (4) ◽

Cited By ~ 18

Author(s):

Maureen J. Donlin ◽

Rajendra Upadhya ◽

Kimberly J. Gerik ◽

Woei Lam ◽

Laura G. VanArendonk ◽

...

Keyword(s):

Cell Wall ◽

Protein Kinase ◽

Cyclic Amp ◽

Protein Kinase A ◽

Cryptococcus Neoformans ◽

Signaling Pathway ◽

Cell Wall Integrity ◽

Wild Type ◽

Content Type ◽

Kinase A

ABSTRACTCryptococcus neoformans is a fungal pathogen of immunocompromised people that causes fatal meningitis. The fungal cell wall is essential to viability and pathogenesis ofC. neoformans, and biosynthesis and repair of the wall is primarily controlled by the cell wall integrity (CWI) signaling pathway. Previous work has shown that deletion of genes encoding the four major kinases in the CWI signaling pathway, namely,PKC1,BCK1,MKK2, andMPK1results in severe cell wall phenotypes, sensitivity to a variety of cell wall stressors, and for Mpk1, reduced virulence in a mouse model. Here, we examined the global transcriptional responses to gene deletions ofBCK1,MKK2, andMPK1compared to wild-type cells. We found that over 1,000 genes were differentially expressed in one or more of the deletion strains, with 115 genes differentially expressed in all three strains, many of which have been identified as genes regulated by the cyclic AMP (cAMP)/protein kinase A (PKA) pathway. Biochemical measurements of cAMP levels in the kinase deletion strains revealed significantly less cAMP in all of the deletion strains compared to the wild-type strain. The deletion strains also produced significantly smaller capsules than the wild-type KN99 strain did under capsule-inducing conditions, although the levels of capsule they shed were similar to those shed by the wild type. Finally, addition of exogenous cAMP led to reduced sensitivity to cell wall stress and restored surface capsule to levels near those of wild type. Thus, we have direct evidence of cross talk between the CWI and cAMP/PKA pathways that may have important implications for regulation of cell wall and capsule homeostasis.IMPORTANCECryptococcus neoformans is a fungal pathogen of immunocompromised people that causes fatal meningitis. The fungal cell wall is essential to viability and pathogenesis ofC. neoformans, and biosynthesis and repair of the wall are primarily controlled by the cell wall integrity (CWI) signaling pathway. In this study, we demonstrate that deletion of any of three core kinases in the CWI pathway impacts not only the cell wall but also the amount of surface capsule. Deletion of any of the kinases results in significantly reduced cellular cyclic AMP (cAMP) levels, and addition of exogenous cAMP rescues the capsule defect and some cell wall defects, supporting a direct role for the CWI pathway in regulation of capsule in conjunction with the cAMP/protein kinase A pathway.

Download Full-text