Pharyngeal polysaccharide deacetylases affect development in the nematode C. elegans and deacetylate chitin in vitro.
Chitin (β-1,4-linked-N-acetylglucosamine) provides structural integrity to the nematode eggshell and pharyngeal lining. Chitin is synthesized in nematodes, but not in plants and vertebrates, which are often hosts to parasitic roundworms; hence, the chitin metabolism pathway is considered a potential target for selective interventions. Polysaccharide deacetylases (PDAs), including those that convert chitin to chitosan, have been previously demonstrated in protists, fungi and insects. We show that genes encoding PDAs are distributed throughout the phylum Nematoda, with the two paralogs F48E3.8 and C54G7.3 found in C. elegans. We confirm that the genes are somatically expressed and show that RNAi knockdown of these genes retards C. elegans development. Additionally, we show that proteins from the nematode deacetylate chitin in vitro, we quantify the substrate available in vivo as targets of these enzymes, and we show that Eosin Y (which specifically stains chitosan in fungal cells walls) stains the C. elegans pharynx. Our results suggest that one function of PDAs in nematodes may be deacetylation of the chitinous pharyngeal lining.
Department of Biology, Tufts University, Medford, Massachusetts, United States of America. ronald.heustis@tufts.edu
Ronald J
RJ
Heustis
Hong K
HK
Ng
Kenneth J
KJ
Brand
Meredith C
MC
Rogers
Linda T
LT
Le
Charles A
CA
Specht
Juliet A
JA
Fuhrman
D000107:Acetylation; D000581:Amidohydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development*; D002686:Chitin; D048271:Chitosan / Q000378:metabolism; D019295:Computational Biology; D018507:Gene Expression Regulation, Developmental; D008969:Molecular Sequence Data; D010614:Pharynx / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development*; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D016415:Sequence Alignment; D012995:Solubility; D013997:Time Factors; D014020:Tissue Extracts
D014020:Tissue Extracts; D002686:Chitin; D048271:Chitosan; D000581:Amidohydrolases; C022427:polysaccharide deacetylase