Sensing stressful conditions that affect the cell wall reorganization is important for yeast survival. Here, we studied two proteins SpWsc1p and SpMtl2p with structural features indicative of plasma membrane-associated cell wall sensors. We found that Mtl2p and Wsc1p act by turning on the Rho1p GTPase. Each gene could be deleted individually without affecting viability, but the deletion of both was lethal and this phenotype was rescued by overexpression of the genes encoding either Rho1p or its GDP/GTP exchange factors (GEFs). In addition, wsc1Δ and mtl2Δ cells showed a low level of Rho1p-GTP under cell wall stress. Mtl2p-GFP (green fluorescent protein) localized to the cell periphery and was necessary for survival under different types of cell wall stress. Wsc1p-GFP was concentrated in patches at the cell tips, it interacted with the Rho-GEF Rgf2p, and its overexpression activated cell wall biosynthesis. Our results are consistent with the notion that cell wall assembly is regulated by two different networks involving Rho1p. One includes signaling from Mtl2p through Rho1p to Pck1p, while the second one implicates signaling from Wsc1p and Rgf2p through Rho1p to activate glucan synthase (GS). Finally, signaling through the mitogen-activated protein kinase (MAPK) Pmk1p remained active in mtl2Δ and wsc1Δ disruptants exposed to cell wall stress, suggesting that the cell wall stress-sensing spectrum of Schizosaccharomyces pombe sensor-like proteins differs from that of Saccharomyces cerevisiae.
It
is known that interactions between wine flavanols and salivary
proline-rich proteins (PRPs) are one of the main factors responsible
for wine astringency. The addition of commercial yeast mannoproteins
(MPs) to wines has been pointed to as a possible tool to modulate
the excessive astringency due to a lack of phenolic maturity at harvest
time that might occur as a consequence of global climate change. The
aim of this work was to study by isothermal titration calorimetry
and molecular dynamics simulation the molecular mechanisms by which
mannoproteins could modulate astringency elicited by tannins and if
it can be influenced by mannoprotein composition. Results obtained
indicate that the MPs assayed had an important impact on astringency
through the formation of ternary aggregates with different solubilities
or by preventing the flavanol–PRP interaction by a competitive
mechanism, although in a different strength, depending on the size
and the compositional characteristic of the mannoprotein.
Fission yeast Rho1p exchange factor Rgf1p is relocalized to the cell nucleus during the stalled replication caused by hydroxyurea. The Cds1p checkpoint kinase, helped by the chaperone Rad24p, controls the nuclear accumulation of Rgf1p by inhibiting its nuclear export. Failure to do so produces inefficient recovery from replication arrest.
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