Assemblies of DivIVA Mark Sites for Hyphal Branching and Can Establish New Zones of Cell Wall Growth in Streptomyces coelicolor

ABSTRACT Time-lapse imaging of Streptomyces hyphae revealed foci of the essential protein DivIVA at sites where lateral branches will emerge. Overexpression experiments showed that DivIVA foci can trigger establishment of new zones of cell wall assembly, suggesting a key role of DivIVA in directing peptidoglycan synthesis and cell shape in Streptomyces.

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The role of the exocytic pathway in cell wall assembly in yeast

Yeast ◽

10.1002/yea.3659 ◽

2021 ◽

Author(s):

Qingguo Guo ◽

Na Meng ◽

Guanzhi Fan ◽

Dong Sun ◽

Yuan Meng ◽

...

Keyword(s):

Cell Wall ◽

Cell Wall Assembly ◽

Wall Assembly ◽

Exocytic Pathway

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Cell Wall Assembly in Saccharomyces cerevisiae

Microbiology and Molecular Biology Reviews ◽

10.1128/mmbr.00038-05 ◽

2006 ◽

Vol 70 (2) ◽

pp. 317-343 ◽

Cited By ~ 489

Author(s):

Guillaume Lesage ◽

Howard Bussey

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Wall ◽

Critical Role ◽

Cell Integrity ◽

Cell Type ◽

Cell Wall Assembly ◽

Regulatory Dynamics ◽

Wall Growth ◽

Secretory System ◽

Wall Assembly

SUMMARY An extracellular matrix composed of a layered meshwork of β-glucans, chitin, and mannoproteins encapsulates cells of the yeast Saccharomyces cerevisiae. This organelle determines cellular morphology and plays a critical role in maintaining cell integrity during cell growth and division, under stress conditions, upon cell fusion in mating, and in the durable ascospore cell wall. Here we assess recent progress in understanding the molecular biology and biochemistry of cell wall synthesis and its remodeling in S. cerevisiae. We then review the regulatory dynamics of cell wall assembly, an area where functional genomics offers new insights into the integration of cell wall growth and morphogenesis with a polarized secretory system that is under cell cycle and cell type program controls.

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The Role of Glucosidase I (Cwh41p) in the Biosynthesis of Cell Wall β-1,6-Glucan Is Indirect

Molecular Biology of the Cell ◽

10.1091/mbc.9.10.2729 ◽

1998 ◽

Vol 9 (10) ◽

pp. 2729-2738 ◽

Cited By ~ 19

Author(s):

Claudia Abeijon ◽

Ling Yun Chen

Keyword(s):

Cell Wall ◽

Double Mutant ◽

Biosynthetic Pathway ◽

Slow Growth ◽

Wild Type ◽

Glucose Residue ◽

Calcofluor White ◽

Cell Wall Assembly ◽

Wall Assembly

CWH41, a gene involved in the assembly of cell wall β-1,6-glucan, has recently been shown to be the structural gene forSaccharomyces cerevisiae glucosidase I that is responsible for initiating the trimming of terminal α-1,2-glucose residue in the N-glycan processing pathway. To distinguish between a direct or indirect role of Cwh41p in the biosynthesis of β-1,6-glucan, we constructed a double mutant, alg5Δ(lacking dolichol-P-glucose synthase) cwh41Δ, and found that it has the same phenotype as the alg5Δsingle mutant. It contains wild-type levels of cell wall β-1,6-glucan, shows moderate underglycosylation of N-linked glycoproteins, and grows at concentrations of Calcofluor White (which interferes with cell wall assembly) that are lethal tocwh41Δ single mutant. The strong genetic interactions of CWH41 with KRE6 andKRE1, two other genes involved in the β-1,6-glucan biosynthetic pathway, disappear in the absence of dolichol-P-glucose synthase (alg5Δ). The triple mutantalg5Δcwh41Δkre6Δ is viable, whereas the double mutant cwh41Δkre6Δ in the same genetic background is not. The severe slow growth phenotype and 75% reduction in cell wall β-1,6-glucan, characteristic of the cwh41Δkre1Δdouble mutant, are not observed in the triple mutantalg5Δcwh41Δkre1Δ. Kre6p, a putative Golgi glucan synthase, is unstable in cwh41Δ strains, and its overexpression renders these cells Calcofluor White resistant. These results demonstrate that the role of glucosidase I (Cwh41p) in the biosynthesis of cell wall β-1,6-glucan is indirect and that dolichol-P-glucose is not an intermediate in this pathway.

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The mycobacterial cell wall partitions the plasma membrane to organize its own synthesis

10.1101/544338 ◽

2019 ◽

Author(s):

Alam García-Heredia ◽

Takehiro Kado ◽

Caralyn E. Sein ◽

Julia Puffal ◽

Sarah H. Osman ◽

...

Keyword(s):

Cell Wall ◽

Plasma Membrane ◽

Membrane Domains ◽

Peptidoglycan Biosynthesis ◽

Cell Wall Assembly ◽

Peptidoglycan Synthesis ◽

Cell Wall Biogenesis ◽

Mycobacterial Cell Wall ◽

Wall Assembly ◽

Made In

AbstractMany antibiotics target the assembly of cell wall peptidoglycan, an essential, heteropolymeric mesh that encases most bacteria. Different species have characteristic subcellular sites of peptidoglycan synthesis that they must carefully maintain for surface integrity and, ultimately, viability. In rod-shaped bacteria, cell wall elongation is spatially precise yet relies on a limited pool of lipid-linked precursors that generate and are attracted to membrane disorder. By tracking enzymes, substrates and products of peptidoglycan biosynthesis in Mycobacterium smegmatis, we show that precursors are made in plasma membrane domains that are laterally and biochemically distinct from sites of cell wall assembly. Membrane partitioning is required for robust, orderly peptidoglycan synthesis, indicating that these domains help template peptidoglycan synthesis. The cell wall-organizing protein DivIVA and the cell wall itself are essential for domain homeostasis. Thus, the peptidoglycan polymer feeds back on its membrane template to maintain an environment conducive to directional synthesis. We further show that our findings are applicable to rod-shaped bacteria that are phylogenetically distant from M. smegmatis, demonstrating that horizontal compartmentalization of precursors is a general feature of bacillary cell wall biogenesis.

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Sophisticated Functions for a Simple Molecule: The Role of Glucosylceramides in Fungal Cells

Lipid Insights ◽

10.4137/lpi.s1014 ◽

2008 ◽

Vol 2 ◽

pp. LPI.S1014 ◽

Cited By ~ 3

Author(s):

Leonardo Nimrichter ◽

Marcio L. Rodrigues ◽

Eliana Barreto-Bergter ◽

Luiz R. Travassos

Keyword(s):

Cell Wall ◽

Immune System ◽

Building Block ◽

Mammalian Cells ◽

Recent Literature ◽

Antifungal Drugs ◽

Cell Wall Assembly ◽

Wall Assembly ◽

Ceramide Moiety

It is well known that mammalian glycosphingolipids (GSL) play key roles in different physiological and pathophysiological processes. The simplest GSL, glucosylceramide (GlcCer), is formed through the enzymatic transfer of glucose to a ceramide moiety. In mammalian cells this molecule is the building block for the synthesis of lactosylceramides and many other complex GSLs. In fungal cells GlcCer is a major neutral GSL that has been considered during decades merely as a structural component of cell membranes. The recent literature, however, describes the participation of fungal GlcCer in vital processes such as secretion, cell wall assembly, recognition by the immune system and regulation of virulence. In this review we discuss the most recent information regarding fungal GlcCer, including (i) new aspects of GlcCer metabolism, (ii) the involvement of these molecules in virulence mechanisms, (iii) their role as targets of new antifungal drugs and immunotherapeutic agents and, finally, (v) their potential participation on cellular signaling in response to different stimuli.

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Modulation of peptidoglycan synthesis by recycled cell wall tetrapeptides

10.1101/771642 ◽

2019 ◽

Cited By ~ 1

Author(s):

Sara B. Hernández ◽

Tobias Dörr ◽

Matthew K. Waldor ◽

Felipe Cava

Keyword(s):

Amino Acids ◽

Cell Wall ◽

Bacterial Cell ◽

Cell Shape ◽

Cell Wall Assembly ◽

Peptidoglycan Synthesis ◽

Cross Linkage ◽

Recycling Pathway ◽

Wall Assembly ◽

Substrate Competition

ABSTRACTThe bacterial cell wall is made of peptidoglycan (PG), a polymer that is essential for maintenance of cell shape and survival. During growth, bacteria remodel their PG, releasing fragments that are predominantly re-internalized by the cell, where they are recycled for synthesis of new PG. Although the PG recycling pathway is widely conserved, its components are not essential and its roles in cell wall homeostasis are not well-understood. Here, we identified LdcV, a Vibrio cholerae L,D-carboxypeptidase that cleaves the terminal D-Alanine from recycled murotetrapeptides. In the absence of ldcV, recycled tetrapeptides accumulated in the cytosol, leading to two toxic consequences for the cell wall. First, incorporation of tetrapeptide-containing PG precursors into the cell wall led to reduction in D,D-cross-linkage between stem peptides, diminishing PG integrity. Second, tetrapeptide accumulation led to a decrease in canonical UDP-pentapeptide precursors, reducing PG synthesis. Thus, LdcV and the recycling pathway promote optimal cell wall assembly and composition. Furthermore, Ldc substrate preference for murotetrapeptides containing canonical (D-Alanine) vs. non-canonical (D-Methionine) D-amino acids is conserved, suggesting that accumulation of tetrapeptide recycling intermediates may modulate PG homeostasis in environments enriched in non-canonical-muropeptides via substrate competition.

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Peptidoglycan precursor synthesis along the sidewall of pole-growing mycobacteria

eLife ◽

10.7554/elife.37243 ◽

2018 ◽

Vol 7 ◽

Cited By ~ 25

Author(s):

Alam García-Heredia ◽

Amol Arunrao Pohane ◽

Emily S Melzer ◽

Caleb R Carr ◽

Taylor J Fiolek ◽

...

Keyword(s):

Cell Wall ◽

Amino Acid ◽

Label Cell ◽

Cell Wall Assembly ◽

Support Cell ◽

Peptidoglycan Synthesis ◽

Precursor Synthesis ◽

Alanine Synthesis ◽

A Site ◽

Wall Assembly

Rod-shaped mycobacteria expand from their poles, yet d-amino acid probes label cell wall peptidoglycan in this genus at both the poles and sidewall. We sought to clarify the metabolic fates of these probes. Monopeptide incorporation was decreased by antibiotics that block peptidoglycan synthesis or l,d-transpeptidation and in an l,d-transpeptidase mutant. Dipeptides complemented defects in d-alanine synthesis or ligation and were present in lipid-linked peptidoglycan precursors. Characterizing probe uptake pathways allowed us to localize peptidoglycan metabolism with precision: monopeptide-marked l,d-transpeptidase remodeling and dipeptide-marked synthesis were coincident with mycomembrane metabolism at the poles, septum and sidewall. Fluorescent pencillin-marked d,d-transpeptidation around the cell perimeter further suggested that the mycobacterial sidewall is a site of cell wall assembly. While polar peptidoglycan synthesis was associated with cell elongation, sidewall synthesis responded to cell wall damage. Peptidoglycan editing along the sidewall may support cell wall robustness in pole-growing mycobacteria.

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Role of turgor pressure in endocytosis in fission yeast

Molecular Biology of the Cell ◽

10.1091/mbc.e13-10-0618 ◽

2014 ◽

Vol 25 (5) ◽

pp. 679-687 ◽

Cited By ~ 55

Author(s):

Roshni Basu ◽

Emilia Laura Munteanu ◽

Fred Chang

Keyword(s):

Cell Wall ◽

Plasma Membrane ◽

Fission Yeast ◽

Turgor Pressure ◽

Time Lapse ◽

Actin Assembly ◽

Latrunculin A ◽

The Media ◽

Time Lapse Imaging

Yeast and other walled cells possess high internal turgor pressure that allows them to grow and survive in the environment. This turgor pressure, however, may oppose the invagination of the plasma membrane needed for endocytosis. Here we study the effects of turgor pressure on endocytosis in the fission yeast Schizosaccharomyces pombe by time-lapse imaging of individual endocytic sites. Decreasing effective turgor pressure by addition of sorbitol to the media significantly accelerates early steps in the endocytic process before actin assembly and membrane ingression but does not affect the velocity or depth of ingression of the endocytic pit in wild-type cells. Sorbitol also rescues endocytic ingression defects of certain endocytic mutants and of cells treated with a low dose of the actin inhibitor latrunculin A. Endocytosis proceeds after removal of the cell wall, suggesting that the cell wall does not contribute mechanically to this process. These studies suggest that endocytosis is governed by a mechanical balance between local actin-dependent inward forces and opposing forces from high internal turgor pressure on the plasma membrane.

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