Expressional and functional variation of horizontally acquired cellulases in the nematode Pristionchus pacificus.
Various whole genome-sequencing projects in the nematode phylum have revealed the widespread occurrence of horizontal gene transfer from different sources. Pristionchus pacificus was the first non-plant parasitic nematode that was found to contain cellulase genes in its genome and to have cellulolytic activity when grown in a monoxenic culture on Escherichia coli. The P. pacificus reference strain PS312 has seven cellulase genes, all of which were acquired by horizontal gene transfer. Previous phylogenomic studies indicated that the acquisition occurred at the base of the genus Pristionchus and was followed by rapid gene duplications and gene turnover. However, little was known about the protein domain architecture, gene expression and the functionality of individual proteins. Here, we analyzed the protein domain architecture, studied the expression at various developmentally stages and tried to induce cellulase gene expression by feeding nematodes with different polysaccharides. Only two of the encoded proteins, Ppa-CEL-2 and Ppa-CEL-3, have a carbohydrate-binding module. Interestingly, these were also the ones with developmental gene expression regulation. Ppa-cel-2 shows high expression in larval and adult stages but is only low expressed in eggs, while Ppa-cel-3 is highly upregulated in adult worms but hardly detectable in any other stage. Ppa-CEL-1, has a catalytic domain similar to Ppa-CEL-2 and Ppa-CEL-3, but lacks a carbohydrate-binding module. The other four cellulases have a very low transcriptional expression correlating with putative incompleteness of their catalytic domain. While, the expression of none of the genes is inducible by polysaccharides, zymographic studies and mass spectrometry indicate that Ppa-CEL-2 and Ppa-CEL-3 are the only two cellulases contributing to cellulase activity in carboxymethylcellulose. Thus, the cellulases of P. pacificus differ in their protein domain architecture, gene expression and functionality. These results indicate that horizontal gene transfer-acquired genes undergo rapid evolutionary changes that affect all aspects of their molecular biology.
Department for Evolutionary Biology, Max Planck Institute for Developmental Biology, Tübingen, Germany.
Lisa N
LN
Schuster
Ralf J
RJ
Sommer
D000818:Animals; D002241:Carbohydrates / Q000737:chemistry; D002266:Carboxymethylcellulose Sodium / Q000737:chemistry; D020134:Catalytic Domain; D002480:Cellulase / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology; D005786:Gene Expression Regulation; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D018014:Gene Transfer Techniques; D022761:Gene Transfer, Horizontal; D023281:Genomics; D006026:Glycoside Hydrolases / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D008957:Models, Genetic; D009348:Nematoda; D010802:Phylogeny; D012313:RNA / Q000378:metabolism
D002241:Carbohydrates; D012313:RNA; D006026:Glycoside Hydrolases; D002480:Cellulase; D002266:Carboxymethylcellulose Sodium