Cellular localization of Sun4p and its interaction with proteins in the yeast birth scar

Kuznetsov, Evgeny; Váchová, Libuše; Palková, Zdena

doi:10.1080/15384101.2016.1189043

Cited by 7 publications

(4 citation statements)

References 24 publications

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“…We focused our search on homologues of SUN4; SUN4 and SIM1 are post-whole genome duplication paralogues, as are UTH1 and NCA3 ( 72 ). These putative glucanases are secreted proteins that localize to the bud scar in S. cerevisiae ( 73 , 74 ). We analysed transcript annotations of SUN4 homologues from other ascomycete fungi C. albicans , N. crassa , A. fumigatus and S. pombe .…”

Section: Resultsmentioning

confidence: 99%

Yeast Ssd1 is a non-enzymatic member of the RNase II family with an alternative RNA recognition site

Bayne

Jayachandran

Kasprowicz

et al. 2021

Nucleic Acids Research

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Ssd1, a conserved fungal RNA-binding protein, is important in stress responses, cell division and virulence. Ssd1 is closely related to Dis3L2 of the RNase II family of nucleases, but lacks catalytic activity and likely suppresses translation of bound mRNAs. Previous studies identified RNA motifs enriched in Ssd1-associated transcripts, yet the sequence requirements for Ssd1 binding are not defined. Here, we identify precise binding sites of Ssd1 on RNA using in vivo cross-linking and cDNA analysis. These sites are enriched in 5′ untranslated regions of a subset of mRNAs encoding cell wall proteins. We identified a conserved bipartite motif that binds Ssd1 with high affinity in vitro. Active RNase II enzymes have a characteristic, internal RNA binding path; the Ssd1 crystal structure at 1.9 Å resolution shows that remnants of regulatory sequences block this path. Instead, RNA binding activity has relocated to a conserved patch on the surface of the protein. Structure-guided mutations of this surface prevent Ssd1 from binding RNA in vitro and phenocopy Ssd1 deletion in vivo. These studies provide a new framework for understanding the function of a pleiotropic post-transcriptional regulator of gene expression and give insights into the evolution of regulatory and binding elements in the RNase II family.

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Section: Resultsmentioning

confidence: 99%

Yeast Ssd1 is a non-enzymatic member of the RNase II family with an alternative RNA recognition site

Bayne

Jayachandran

Kasprowicz

et al. 2021

Nucleic Acids Research

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show abstract

“…We focused our search on homologues of SUN4; SUN4 and SIM1 are post-whole genome duplication paralogues, as are UTH1 and NCA3 (Byrne and Wolfe 2005). These putative glucanases are secreted proteins that localise to the bud scar in S. cerevisiae (Kuznetsov et al 2013; Kuznetsov, Vachova, and Palkova 2016). We analysed transcript annotations of SUN4 homologues from other ascomycete fungi Candida albicans, Neurospora crassa, Aspergillus fumigatus , and S. pombe .…”

Section: Resultsmentioning

confidence: 99%

Yeast Ssd1 is a non-enzymatic member of the RNase II family with an alternative RNA recognition interface

Bayne

Jayachandran

Kasprowicz

et al. 2020

Preprint

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The conserved fungal RNA binding protein Ssd1, is important in stress responses, cell division and virulence. Ssd1 is closely related to Dis3L2 of the RNase II family of nucleases, but lacks catalytic activity and may act by suppressing translation of associated mRNAs. Previous studies identified motifs that are enriched in Ssd1-associated transcripts, yet the sequence requirements for Ssd1 binding are not well understood. Here we present the crystal structure of Ssd1 at 1.9 Å resolution. Active RNase II enzymes have a characteristic, internal RNA binding path, but in Ssd1 this is blocked by remnants of regulatory sequences. Instead, RNA binding activity has likely been relocated to the outer surface of the protein. Using in vivo crosslinking and cDNA analysis (CRAC), we identify Ssd1-RNA binding sites. These are strongly enriched in 5′UTRs of a subset of mRNAs encoding cell wall proteins. Based on these and previous analyses, we identified a conserved bipartite motif that binds Ssd1 with high affinity in vitro. These studies provide a new framework for understanding the function of a pleiotropic post-transcriptional regulator of gene expression and give insights into the evolution of regulatory elements in the RNase II family.

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“…Other cluster V member genes ( EGT2 and SUN4 ) have also been reported to encode glycosidases, and our results clearly showed that genetic perturbation prevented efficient daughter cell separation in all cluster V mutants. Consistent with the mutant phenotypes, co-localization studies have revealed that Dse2, Egt2, and Sun4 form a complex at the birth scar [ 51 ]. Cluster V mutants exhibited no defects in cell-cycle progression or daughter cell growth in the next cell cycle, indicating lesser effects of these genes in cell proliferation.…”

Section: Discussionmentioning

confidence: 87%

Defining Functions of Mannoproteins in Saccharomyces cerevisiae by High-Dimensional Morphological Phenotyping

Ghanegolmohammadi

Okada

Liu

et al. 2021

JoF

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Mannoproteins are non-filamentous glycoproteins localized to the outermost layer of the yeast cell wall. The physiological roles of these structural components have not been completely elucidated due to the limited availability of appropriate tools. As the perturbation of mannoproteins may affect cell morphology, we investigated mannoprotein mutants in Saccharomyces cerevisiae via high-dimensional morphological phenotyping. The mannoprotein mutants were morphologically classified into seven groups using clustering analysis with Gaussian mixture modeling. The pleiotropic phenotypes of cluster I mutant cells (ccw12Δ) indicated that CCW12 plays major roles in cell wall organization. Cluster II (ccw14Δ, flo11Δ, srl1Δ, and tir3Δ) mutants exhibited altered mother cell size and shape. Mutants of cluster III and IV exhibited no or very small morphological defects. Cluster V (dse2Δ, egt2Δ, and sun4Δ) consisted of endoglucanase mutants with cell separation defects due to incomplete septum digestion. The cluster VI mutant cells (ecm33Δ) exhibited perturbation of apical bud growth. Cluster VII mutant cells (sag1Δ) exhibited differences in cell size and actin organization. Biochemical assays further confirmed the observed morphological defects. Further investigations based on various omics data indicated that morphological phenotyping is a complementary tool that can help with gaining a deeper understanding of the functions of mannoproteins.

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Cellular localization of Sun4p and its interaction with proteins in the yeast birth scar

Cited by 7 publications

References 24 publications

Yeast Ssd1 is a non-enzymatic member of the RNase II family with an alternative RNA recognition site

Yeast Ssd1 is a non-enzymatic member of the RNase II family with an alternative RNA recognition site

Yeast Ssd1 is a non-enzymatic member of the RNase II family with an alternative RNA recognition interface

Defining Functions of Mannoproteins in Saccharomyces cerevisiae by High-Dimensional Morphological Phenotyping

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