Global mRNA expression analysis in myosin II deficient strains of Saccharomyces cerevisiae reveals an impairment of cell integrity functions

Rodríguez-Quiñones, José F; Irizarry, Rafael A.; Díaz-Blanco, Nitza L.; Rivera‐Molina, Felix; Gómez-Garzón, Diana; Rodríguez‐Medina, José R.

doi:10.1186/1471-2164-9-34

Cited by 12 publications

(41 citation statements)

References 27 publications

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“…Of 137 RP genes in the yeast genome, 46 were down regulated post-transcriptionally in the myo1Δ strain with 83 previously shown to be down regulated by global analysis and 8 reflecting no change in their steady state or translation levels [6]. These results indicate that in addition to the reported transcriptional regulation of RP genes, there can be significant co-regulation of these genes (35/137) in myo1Δ strains.…”

Section: Discussionmentioning

confidence: 68%

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Post-transcriptional regulation in the myo1Δ mutant of Saccharomyces cerevisiae

et al. 2010

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BackgroundSaccharomyces cerevisiae myosin type II-deficient (myo1Δ) strains remain viable and divide, despite the absence of a cytokinetic ring, by activation of the PKC1-dependent cell wall integrity pathway (CWIP). Since the myo1Δ transcriptional fingerprint is a subset of the CWIP fingerprint, the myo1Δ strain may provide a simplified paradigm for cell wall stress survival.ResultsTo explore the post-transcriptional regulation of the myo1Δ stress response, 1,301 differentially regulated ribosome-bound mRNAs were identified by microarray analysis of which 204 were co-regulated by transcription and translation. Four categories of mRNA were significantly affected - protein biosynthesis, metabolism, carbohydrate metabolism, and unknown functions. Nine genes of the 20 CWIP fingerprint genes were post-transcriptionally regulated. Down and up regulation of selected ribosomal protein and cell wall biosynthesis mRNAs was validated by their distribution in polysomes from wild type and myo1Δ strains. Western blot analysis revealed accumulation of the phosphorylated form of eukaryotic translation initiation factor 2 (eIF2α-P) and a reduction in the steady state levels of the translation initiation factor eIF4Gp in myo1Δ strains. Deletion of GCN2 in myo1Δ abolished eIF2αp phosphorylation, and showed a severe growth defect. The presence of P-bodies in myo1Δ strains suggests that the process of mRNA sequestration is active, however, the three representative down regulated RP mRNAs, RPS8A, RPL3 and RPL7B were present at equivalent levels in Dcp2p-mCh-positive immunoprecipitated fractions from myo1Δ and wild type cells. These same RP mRNAs were also selectively co-precipitated with eIF2α-P in myo1Δ strains.ConclusionsQuantitative analysis of ribosome-associated mRNAs and their polyribosome distributions suggests selective regulation of mRNA translation efficiency in myo1Δ strains. Inhibition of translation initiation factor eIF2α (eIF2α-P) in these strains was by Gcn2p-dependent phosphorylation. The increase in the levels of eIF2α-P; the genetic interaction between GCN2 and MYO1; and the reduced levels of eIF4Gp suggest that other signaling pathways, in addition to the CWIP, may be important for myo1Δ strain survival. Selective co-immunoprecipitation of RP mRNAs with eIF2α-P in myo1Δ strains suggests a novel mode of translational regulation. These results indicate that post-transcriptional control is important in the myo1Δ stress response and possibly other stresses in yeast.

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Section: Discussionmentioning

confidence: 68%

“…Experiments were performed as described in [6], 1.0 μg of total RNA extracted from ribosomal pellets was amplified using the Low RNA Input Fluorescent Linear Amplification kit (Agilent Technologies, Palo Alto, CA). Amplified cRNA was labeled with 10 mM Cyanine 5-CTP (Cy5) or Cyanine 3-CTP (Cy3) (Perkin Elmer Life Sciences, Boston, MA).…”

Section: Methodsmentioning

confidence: 99%

Post-transcriptional regulation in the myo1Δ mutant of Saccharomyces cerevisiae

et al. 2010

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“…During SS formation, Rho1 effectors implicated in cell wall synthesis and stress response are likely to be activated, such as the glucan synthases Fks1 and Fks2 (Mazur and Baginsky, 1996;Qadota et al, 1996), the protein kinase Pkc1 (through activation of the MAPK pathway, in which Kdx1 [also known as Mlp1] is involved; Watanabe et al, 1997;Kim et al, 2008), and Chs3 (Valdivia and Schekman, 2003;Yoshida et al, 2009). In support of this hypothesis, deletions of these genes exhibit synthetic growth defects with myo1 (our unpublished data; Rodríguez-Quiñones et al, 2008;Yoshida et al, 2009).…”

Section: Antagonistic Regulation Of Ss Formation By Rho1 and Cdc42mentioning

confidence: 63%

Distinct roles of Rho1, Cdc42, and Cyk3 in septum formation and abscission during yeast cytokinesis

Onishi

Nishihama

et al. 2013

Journal of Cell Biology

109

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“…As reference genes, we applied ACT1 encoding actin, traditionally used in expression studies of S. cerevisiae by Northern blot or quantitative RT-PCR (e.g. Rodríguez-Quiñones et al 2008) and MGE1 encoding mitochondrial GrpE protein which expression remains unchanged over a wide range of redox conditions (Budzinska et al 2007;Galganska et al 2008). These genes belong to different functional categories that minimizes the risk of coregulation.…”

Section: Resultsmentioning

confidence: 99%

VDAC contributes to mRNA levels in Saccharomyces cerevisiae cells by the intracellular reduction/oxidation state dependent and independent mechanisms

Gałgańska

Antoniewicz

Budzińska

et al. 2010

J Bioenerg Biomembr

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Available data suggest that voltage-dependent anion selective channel (VDAC) constitutes an important component of a cellular regulatory mechanism based on the intracellular reduction/oxidation (redox) state. Here, using quantitative RT-PCR, we demonstrated that depletion of VDAC1 (termed here VDAC) in Saccharomyces cerevisiae cells distinctly affected levels of mRNAs encoding nuclear proteins sensitive to changes of the intracellular redox state including the nuclear transcription factors important for adaptation to the redox state and proteins involved in communication between mitochondria and the nucleus. We also revealed that the changes of the studied protein transcript levels generally correlated with changes of the intracellular redox state although VDAC appears also to affect mRNA levels by a mechanism not based on changes of the intracellular redox states. Thus, VDAC seems to be an important element of the intracellular signaling network.

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Global mRNA expression analysis in myosin II deficient strains of Saccharomyces cerevisiae reveals an impairment of cell integrity functions

Cited by 12 publications

References 27 publications

Post-transcriptional regulation in the myo1Δ mutant of Saccharomyces cerevisiae

Post-transcriptional regulation in the myo1Δ mutant of Saccharomyces cerevisiae

Distinct roles of Rho1, Cdc42, and Cyk3 in septum formation and abscission during yeast cytokinesis

VDAC contributes to mRNA levels in Saccharomyces cerevisiae cells by the intracellular reduction/oxidation state dependent and independent mechanisms

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