Phosphorylation of mRNA-Binding Proteins Puf1 and Puf2 by TORC2-Activated Protein Kinase Ypk1 Alleviates Their Repressive Effects

Galez, Henri A.; Roelants, Françoise M.; Palm, Sarah M.; Reynaud, Kendra; Ingolia, Nicholas T.; Thorner, Jeremy

doi:10.3390/membranes11070500

Cited by 4 publications

(7 citation statements)

References 90 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Puf2p is a member of the Puf family regulator that enables mRNA binding. Puf2p has shown a preference for interaction with mRNAs encoding plasma membrane-associated proteins to regulate the stability, localization, and efficiency of translation of the bound transcript [ 15 , 16 ]. GPI12 encodes an N-acetylglucosaminyl phosphatidylinositol deacetylase located in the endoplasmic reticulum, which was shown to catalyze the deacetylation of N-acetylglucosaminyl phosphatidylinositol to produce glucosaminyl phosphatidylinositol [ 17 ].…”

Section: Resultsmentioning

confidence: 99%

Evolutionary and reverse engineering in Saccharomyces cerevisiae reveals a Pdr1p mutation-dependent mechanism for 2-phenylethanol tolerance

Xia

Kang

et al. 2022

Microb Cell Fact

View full text Add to dashboard Cite

Background 2-Phenylethanol (2-PE), a higher alcohol with a rose-like odor, inhibits growth of the producer strains. However, the limited knowledge regarding 2-PE tolerance mechanisms renders our current knowledge base insufficient to inform rational design. Results To improve the growth phenotype of Saccharomyces cerevisiae under a high 2-PE concentration, adaptive laboratory evolution (ALE) was used to generate an evolved 19–2 strain. Under 2-PE stress, its OD600 and growth rate increased by 86% and 22% than that of the parental strain, respectively. Through whole genome sequencing and reverse engineering, transcription factor Pdr1p mutation (C862R) was revealed as one of the main causes for increased 2-PE tolerance. Under 2-PE stress condition, Pdr1p mutation increased unsaturated fatty acid/saturated fatty acid ratio by 42%, and decreased cell membrane damage by 81%. Using STRING website, we identified Pdr1p interacted with some proteins, which were associated with intracellular ergosterol content, reactive oxygen species (ROS), and the ATP-binding cassette transporter. Also, the results of transcriptional analysis of genes encoded these proteins confirmed that Pdr1p mutation induced the expression of these genes. Compared with those of the reference strain, the ergosterol content of the PDR1_862 strain increased by 72%–101%, and the intracellular ROS concentration decreased by 38% under 2-PE stress. Furthermore, the Pdr1p mutation also increased the production of 2-PE (11% higher). Conclusions In the present work, we have demonstrated the use of ALE as a powerful tool to improve yeast tolerance to 2-PE. Based on the reverse engineering, transcriptional and physiological analysis, we concluded that Pdr1p mutation significantly enhanced the 2-PE tolerance of yeast by regulating the fatty acid proportion, intracellular ergosterol and ROS. It provides new insights on Pdr1p mediated 2-PE tolerance, which could help in the design of more robust yeasts for natural 2-PE synthesis.

show abstract

Section: Resultsmentioning

confidence: 99%

Evolutionary and reverse engineering in Saccharomyces cerevisiae reveals a Pdr1p mutation-dependent mechanism for 2-phenylethanol tolerance

Xia

Kang

et al. 2022

Microb Cell Fact

View full text Add to dashboard Cite

show abstract

“…Remarkably, consistent with all of the studies discussed above documenting that a primary physiological role of TORC2-Ypk1 signaling in yeast is homeostatic maintenance of the components of the plasma membrane, prior work by others had demonstrated that Puf1 and Puf2 exhibit a marked preference for interaction with mRNAs encoding plasma membrane-associated proteins, including integral membrane proteins [ 226 , 227 ]. Indeed, we were able to show, first, that both Puf1 and Puf2 are authentic Ypk1 substrates both in vitro and in vivo [ 228 ]. Second, we found that fluorescently tagged Puf1 localizes constitutively in cortical puncta closely apposed to the plasma membrane, whereas Puf2 does so in the absence of its Ypk1 phosphorylation, but is dispersed in the cytosol when phosphorylated [ 228 ].…”

Section: Functions Of Ypk1mentioning

confidence: 99%

“…Indeed, we were able to show, first, that both Puf1 and Puf2 are authentic Ypk1 substrates both in vitro and in vivo [ 228 ]. Second, we found that fluorescently tagged Puf1 localizes constitutively in cortical puncta closely apposed to the plasma membrane, whereas Puf2 does so in the absence of its Ypk1 phosphorylation, but is dispersed in the cytosol when phosphorylated [ 228 ]. We further demonstrated that Ypk1-mediated phosphorylation of Puf1 and Puf2 up-regulates the production of the protein products of the transcripts to which they bind, with a concomitant increase in the level of the cognate mRNAs [ 228 ].…”

Section: Functions Of Ypk1mentioning

confidence: 99%

“…Second, we found that fluorescently tagged Puf1 localizes constitutively in cortical puncta closely apposed to the plasma membrane, whereas Puf2 does so in the absence of its Ypk1 phosphorylation, but is dispersed in the cytosol when phosphorylated [ 228 ]. We further demonstrated that Ypk1-mediated phosphorylation of Puf1 and Puf2 up-regulates the production of the protein products of the transcripts to which they bind, with a concomitant increase in the level of the cognate mRNAs [ 228 ]. Thus, Ypk1 phosphorylation relieves Puf1- and Puf2-mediated post-transcriptional repression mainly by counteracting their negative effect on transcript stability.…”

Section: Functions Of Ypk1mentioning

confidence: 99%

“…Thus, Ypk1 phosphorylation relieves Puf1- and Puf2-mediated post-transcriptional repression mainly by counteracting their negative effect on transcript stability. Moreover, using a heterologous protein–RNA tethering and fluorescent protein reporter assay, the consequence of Ypk1 phosphorylation in vivo was recapitulated for full-length Puf1 and even for N-terminal fragments (residues 1–340 and 143–295) corresponding to the region upstream of its dimerization domain (an RNA-recognition motif fold) encompassing its two Ypk1 phosphorylation sites (both also conserved in Puf2) [ 228 ]. This latter result suggests that alleviation of Puf1-imposed transcript destabilization does not obligatorily require dissociation of Ypk1-phosphorylated Puf1 from a transcript.…”

Section: Functions Of Ypk1mentioning

confidence: 99%

See 2 more Smart Citations

TOR complex 2 is a master regulator of plasma membrane homeostasis

Thorner

2022

Biochemical Journal

Self Cite

View full text Add to dashboard Cite

As first demonstrated in budding yeast (Saccharomyces cerevisiae), all eukaryotic cells contain two, distinct multi-component protein kinase complexes that each harbor the TOR (Target Of Rapamycin) polypeptide as the catalytic subunit. These ensembles, dubbed TORC1 and TORC2, function as universal, centrally important sensors, integrators, and controllers of eukaryotic cell growth and homeostasis. TORC1, activated on the cytosolic surface of the lysosome (or, in yeast, on the cytosolic surface of the vacuole), has emerged as a primary nutrient sensor that promotes cellular biosynthesis and suppresses autophagy. TORC2, located primarily at the plasma membrane, plays a major role in maintaining the proper levels and bilayer distribution of all plasma membrane components (sphingolipids, glycerophospholipids, sterols, and integral membrane proteins). This article surveys what we have learned about signaling via the TORC2 complex, largely through studies conducted in S. cerevisiae. In this yeast, conditions that challenge plasma membrane integrity can, depending on the nature of the stress, stimulate or inhibit TORC2, resulting in, respectively, up-regulation or down-regulation of the phosphorylation and thus the activity of its essential downstream effector the AGC family protein kinase Ypk1. Through the ensuing effect on the efficiency with which Ypk1 phosphorylates multiple substrates that control diverse processes, membrane homeostasis is maintained. Thus, the major focus here is on TORC2, Ypk1, and the multifarious targets of Ypk1 and how the functions of these substrates are regulated by their Ypk1-mediated phosphorylation, with emphasis on recent advances in our understanding of these processes.

show abstract

Stop, slow-down or speed-up? : The role of mRNA binding proteins during seed germination

Sajeev

View full text Add to dashboard Cite

The seed life cycle Seed development and maturation: The birth of a seedArabidopsis seed development is composed of two phases, namely, the embryogenesis and the seed maturation phase which partially overlap with each other (Fig. 1.1) (Bewley et al., 2012). The embryogenesis phase, starts with a double fertilization event that leads to a single zygote. This zygotic embryo grows filling the embryo sac through cell division and expansion, and ends with all the embryo structures being formed. At the end of this phase, cell division is arrested (Fig. 1.1A) (Raz et al., 2001). While the embryo is still transforming morphologically, the first part of the maturation phase commences (Raz et al., 2001;Nonogaki, 2019). The maturation phase begins with accumulation of reserves like storage proteins, mRNAs and sugars important for the germination process (Fig. 1.1B). This is followed by an intense period of moisture loss and ends with the embryo being in a quiescent and dry state. In this dehydrated state, desiccation tolerance and primary dormancy have been established (Fig 1 .1C) (Gutierrez et al., 2007;Holdsworth et al., 2008). The mature seed houses an embryo that forms the new plant, a single layer of surrounding tissue often referred to as the endosperm, and the seed coat (testa), a protective outer layer of dead tissue (

show abstract

Phosphorylation of mRNA-Binding Proteins Puf1 and Puf2 by TORC2-Activated Protein Kinase Ypk1 Alleviates Their Repressive Effects

Cited by 4 publications

References 90 publications

Evolutionary and reverse engineering in Saccharomyces cerevisiae reveals a Pdr1p mutation-dependent mechanism for 2-phenylethanol tolerance

Evolutionary and reverse engineering in Saccharomyces cerevisiae reveals a Pdr1p mutation-dependent mechanism for 2-phenylethanol tolerance

TOR complex 2 is a master regulator of plasma membrane homeostasis

Stop, slow-down or speed-up? : The role of mRNA binding proteins during seed germination

Contact Info

Product

Resources

About