Characterization of the Caenorhabditis elegans UDP-galactopyranose mutase homolog glf-1 reveals an essential role for galactofuranose metabolism in nematode surface coat synthesis

Novelli, Jacopo; Chaudhary, Kshitiz; Canovas, Julie; Benner, Jack S.; Madinger, Catherine L.; Kelly, Paul A.; Hodgkin, Jonathan; Carlow, Clotilde K. S.

doi:10.1016/j.ydbio.2009.09.010

Cited by 38 publications

(42 citation statements)

References 94 publications

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“…NA, not applicable. (Partridge et al 2008), and bus-17 (K. J. Yook and C. Darby, unpublished observations), as well as for glf-1 (Novelli et al 2009). These observations are in accord with the belief that the seam cells, rather than the hypodermis, are responsible for secreting the surface coat.…”

Section: Discussionmentioning

confidence: 91%

“…Both viable and lethal alleles of bus-8 have been identified; weak mutants of bus-8 such as e2698 are viable and resistant to M. nematophilum (Bus phenotype), but they still permit formation of Yersinia biofilms, in contrast to other mutants of this type. Another essential carbohydrate-modifying gene required for cuticle integrity is glf-1, encoding UDP-galactopyranose mutase (Novelli et al 2009). Available mutants in this gene are barely viable, making it difficult to assess interactions with M. nematophilum or Yersinia, but a glf-1 hypomorph (made by partial rescue of a lethal glf-1 mutant with a Leishmania-derived transgene) is resistant to M. nematophilum infection, indicating that this gene, like bus-8, is essential both for surface barrier formation and for bacterial surface infection.…”

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confidence: 99%

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Glycosylation Genes Expressed in Seam Cells Determine Complex Surface Properties and Bacterial Adhesion to the Cuticle of Caenorhabditis elegans

et al. 2011

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The surface of the nematode Caenorhabditis elegans is poorly understood but critical for its interactions with the environment and with pathogens. We show here that six genes (bus-2, bus-4, and bus-12, together with the previously cloned srf-3, bus-8, and bus-17) encode proteins predicted to act in surface glycosylation, thereby affecting disease susceptibility, locomotory competence, and sexual recognition. Mutations in all six genes cause resistance to the bacterial pathogen Microbacterium nematophilum, and most of these mutations also affect bacterial adhesion and biofilm formation by Yersinia species, demonstrating that both infection and biofilm formation depend on interaction with complex surface carbohydrates. A new bacterial interaction, involving locomotory inhibition by a strain of Bacillus pumilus, reveals diversity in the surface properties of these mutants. Another biological property-contact recognition of hermaphrodites by males during mating-was also found to be impaired in mutants of all six genes. An important common feature is that all are expressed most strongly in seam cells, rather than in the main hypodermal syncytium, indicating that seam cells play the major role in secreting surface coat and consequently in determining environmental interactions. To test for possible redundancies in gene action, the 15 double mutants for this set of genes were constructed and examined, but no synthetic phenotypes were observed. Comparison of the six genes shows that each has distinctive properties, suggesting that they do not act in a linear pathway.

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Section: Discussionmentioning

confidence: 91%

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confidence: 99%

Glycosylation Genes Expressed in Seam Cells Determine Complex Surface Properties and Bacterial Adhesion to the Cuticle of Caenorhabditis elegans

et al. 2011

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“…The need for substantially more compound to detect activity against the nematode has also been reported in other enzyme-based screening campaigns for new filarial leads [41]–[43]. It is thought that the outermost layer of the cuticle is the main barrier to penetration of drugs, stains, and other chemicals [44], [45]. Further studies are needed to determine the physicochemical factors that favor the absorption of compounds in nematodes.…”

Section: Discussionmentioning

confidence: 98%

A Target Repurposing Approach Identifies N-myristoyltransferase as a New Candidate Drug Target in Filarial Nematodes

Galvin

Villemaine

et al. 2014

PLoS Negl Trop Dis

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Myristoylation is a lipid modification involving the addition of a 14-carbon unsaturated fatty acid, myristic acid, to the N-terminal glycine of a subset of proteins, a modification that promotes their binding to cell membranes for varied biological functions. The process is catalyzed by myristoyl-CoA:protein N-myristoyltransferase (NMT), an enzyme which has been validated as a drug target in human cancers, and for infectious diseases caused by fungi, viruses and protozoan parasites. We purified Caenorhabditis elegans and Brugia malayi NMTs as active recombinant proteins and carried out kinetic analyses with their essential fatty acid donor, myristoyl-CoA and peptide substrates. Biochemical and structural analyses both revealed that the nematode enzymes are canonical NMTs, sharing a high degree of conservation with protozoan NMT enzymes. Inhibitory compounds that target NMT in protozoan species inhibited the nematode NMTs with IC50 values of 2.5–10 nM, and were active against B. malayi microfilariae and adult worms at 12.5 µM and 50 µM respectively, and C. elegans (25 µM) in culture. RNA interference and gene deletion in C. elegans further showed that NMT is essential for nematode viability. The effects observed are likely due to disruption of the function of several downstream target proteins. Potential substrates of NMT in B. malayi are predicted using bioinformatic analysis. Our genetic and chemical studies highlight the importance of myristoylation in the synthesis of functional proteins in nematodes and have shown for the first time that NMT is required for viability in parasitic nematodes. These results suggest that targeting NMT could be a valid approach for the development of chemotherapeutic agents against nematode diseases including filariasis.

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“…14 for a recent review). Genetic and biochemical studies have shown that UDP-Galf arising from the action of UGM is essential for galactofuranosylation in both prokaryotes and eukaryotes (2,12,13,15,16). This nucleotide sugar is the established or presumed donor substrate of specific galactofuranosyltransferases (GlfTs) comprising characterized bacterial ␤-GlfTs (14) and a family of putative ␤-GlfTs identified in the protozoan parasite L. major (17,18).…”

Section: In Vitro Transport Assays Established Binding Of Udp-galf Tomentioning

confidence: 99%

A Single UDP-galactofuranose Transporter Is Required for Galactofuranosylation in Aspergillus fumigatus

Engel

Schmalhorst

Dörk

et al. 2009

Journal of Biological Chemistry

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Galactofuranose (Galf) containing molecules have been described at the cell surface of several eukaryotes and shown to contribute to the virulence of the parasite Leishmania major and the fungus Aspergillus fumigatus. It is anticipated that a number of the surface glycoconjugates such as N-glycans or glycolipids are galactofuranosylated in the Golgi apparatus. This raises the question of how the substrate for galactofuranosylation reactions, UDP-Galf, which is synthesized in the cytosol, translocates into the organelles of the secretory pathway. Here we report the first identification of a Golgi-localized nucleotide sugar transporter, named GlfB, with specificity for a UDP-Galf. In vitro transport assays established binding of UDP-Galf toGlfB and excluded transport of several other nucleotide sugars. Furthermore, the implication of glfB in the galactofuranosylation of A. fumigatus glycoconjugates and galactomannan was demonstrated by a targeted gene deletion approach. Our data reveal a direct connection between galactomannan and the organelles of the secretory pathway that strongly suggests that the cell wall-bound polysaccharide originates from its glycosylphosphatidylinositol-anchored form.

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Characterization of the Caenorhabditis elegans UDP-galactopyranose mutase homolog glf-1 reveals an essential role for galactofuranose metabolism in nematode surface coat synthesis

Cited by 38 publications

References 94 publications

Glycosylation Genes Expressed in Seam Cells Determine Complex Surface Properties and Bacterial Adhesion to the Cuticle of Caenorhabditis elegans

Glycosylation Genes Expressed in Seam Cells Determine Complex Surface Properties and Bacterial Adhesion to the Cuticle of Caenorhabditis elegans

A Target Repurposing Approach Identifies N-myristoyltransferase as a New Candidate Drug Target in Filarial Nematodes

A Single UDP-galactofuranose Transporter Is Required for Galactofuranosylation in Aspergillus fumigatus

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