Major sperm protein genes from Onchocerca volvulus

Molecular and Biochemical Parasitology, 2005

Nematode sperm utilize a crawling motility based on a nematode sperm-specific cytoskeletal protein called the major sperm protein (MSP). Although MSP has no similarity to actin in sequence or structure, the motility mediated by these two proteins is nearly indistinguishable at a phenotypic level. As with the traditional actin cytoskeleton, the central component of MSP-based motility (MSP) interacts with accessory proteins that regulate polymerization and depolymerization and play a key role in cell motility. A bioinformatics approach has led to the identification of proteins with enhanced expression in the Ascaris suum male germ line, including five new proteins each containing an MSP domain. One of these MSP domain proteins (As-MDP-1) contains an MSP domain in the C-terminus and a N-terminal extension rich in prolines and alanines. The 15.6 kDa As-MDP-1 was shown to be >90% identical at the amino acid level to members of a small family of Ascaris proteins that have been shown to bind to the MSP cytoskeleton and to negatively regulate MSP fiber growth. Further, it was demonstrated that As-MDP-1 is the smallest member of the MFP1 triplet of negative regulators of MSP cytoskeleton formation. Antibodies were used to detect the presence of As-MDP-1 along the entire length of the MSP fibers suggesting that As-MDP-1 binds directly to the higher order forms of MSP. This protein has orthologues in other nematode species, is present in Ascaris in at least six allelic forms, and is likely to form multimers. stranded ␤ sheets [3]. The MSPs from A. suum and from the free-living nematode species Caenorhabditis elegans form homodimers in solution which in turn polymerize into an ordered series of helical structures . MSP dimers polymerize into extended filaments that are composed of two helical strands. The filaments of MSP are anchored to the pseudopod membrane at the advancing front of the cell where they are cross-linked into interconnecting fiber complexes. The fiber complexes flow rearward where they disassemble so that core constituents can be recycled to the leading edge for reassembly . The assembly and disassembly of the MSP cytoskeleton is mediated in part by regional differences in intracellular pH .

Molecular and Cellular Biology, 1984

2005

the many collaborators who have generously provided materials (www.nematode.net/Collaborators/), especially Thomas Baum for supplying staged libraries of Heterodera glycines. JPM and DMB are equity holders of Divergence Inc., and JPM is a Divergence employee; this research was not company funded.

Journal of …, 2003

the many collaborators who have generously provided materials (www.nematode.net/Collaborators/), especially Thomas Baum for supplying staged libraries of Heterodera glycines. JPM and DMB are equity holders of Divergence Inc., and JPM is a Divergence employee; this research was not company funded.

2014

The organization of genes into operons, clusters of genes that are co-transcribed to produce polycistronic pre-mRNAs, is a trait found in a wide range of eukaryotic groups, including multiple animal phyla. Operons are present in the class Chromadorea, one of the two main nematode classes, but their distribution in the other class, the Enoplea, is not known. We have surveyed the genomes of Trichinella spiralis, Trichuris muris, and Romanomermis culicivorax and identified the first putative operons in members of the Enoplea. Consistent with the mechanism of polycistronic RNA resolution in other nematodes, the mRNAs produced by genes downstream of the first gene in the T. spiralis and T. muris operons are trans-spliced to spliced leader RNAs, and we are able to detect polycistronic RNAs derived from these operons. Importantly, a putative intercistronic region from one of these potential enoplean operons confers polycistronic processing activity when expressed as part of a chimeric operon in Caenorhabditis elegans. We find that T. spiralis genes located in operons have an increased likelihood of having operonic C. elegans homologs. However, operon structure in terms of synteny and gene content is not tightly conserved between the two taxa, consistent with models of operon evolution. We have nevertheless identified putative operons conserved between Enoplea and Chromadorea. Our data suggest that operons and "spliced leader" (SL) trans-splicing predate the radiation of the nematode phylum, an inference which is supported by the phylogenetic profile of proteins known to be involved in nematode SL trans-splicing.

eLS, 2001

In the past few years, an increasing number of draft genome sequences of multiple free-living and parasitic nematodes have been published. Although nematode genomes vary in size within an order of magnitude, compared with mammalian genomes, they are all very small. Nevertheless, nematodes possess only marginally fewer genes than mammals do. Nematode genomes are very compact and therefore form a highly attractive system for comparative studies of genome structure and evolution. Strikingly, approximately one-third of the genes in every sequenced nematode genome has no recognisable homologues outside their genus. One observes high rates of gene losses and gains, among them numerous examples of gene acquisition by horizontal gene transfer. Not only does the 'gene for parasitism' not exist, but also there appear to be no common genomic characteristics of parasitic nematode genomes which would distinguish them from genomes of free-living nematodes.

The parasitic nematode intestine is responsible for nutrient digestion and absorption, and many other processes essential for reproduction and survival, making it a valuable target for anthelmintic drug treatment. However, nematodes display extreme biological diversity (including occupying distinct trophic habitats), resulting in limited knowledge of intestinal cell/protein functions of fundamental or adaptive significance. We developed a perfusion model for isolating intestinal proteins in Ascaris suum (a parasite of humans and swine), allowing for the identification of over 1000 intestinal A. suum proteins (using mass spectrometry), which were assigned to several different intestinal cell compartments (intestinal tissue, the integral and peripheral intestinal membranes, and the intestinal lumen). A multi-omics analysis approach identified a large diversity of biological functions across intestinal compartments, based on both functional enrichment analysis (identifying terms related to detoxification, proteolysis, and host-parasite interactions) and regulatory binding sequence analysis to identify putatively active compartment-specific transcription factors (identifying many related to intestinal sex differentiation or lifespan regulation). Orthologs of A. suum proteins in 15 other nematodes species, five host species, and two outgroups were identified and analyzed. Different cellular compartments demonstrated markedly different levels of protein conservation; e.g. integral intestinal membrane proteins were the most conserved among nematodes (up to 96% conservation), whereas intestinal lumen proteins were the most diverse (only 6% conservation across all nematodes, and 71% with no host orthologs). Finally, this integrated multi-omics analysis identified conserved nematode-specific intestinal proteins likely performing essential functions (including V-type ATPases and ABC transporters), which may serve as promising anthelmintic drug or vaccine targets in future research. Collectively, the findings provide valuable new insights on conserved and adaptive features of nematode intestinal cells, membranes and the intestinal lumen, and potential targets for parasite treatment and control. Molecular & Cellular

Parasitology Research, 2008

Populations of the bovine lungworm, Dictyocaulus viviparus, are genetically structured based on variation in mtDNA and AFLP data. Our aim was to investigate if this genetic variability also is reflected in a protein recognized by the host immune system. We focused on the major sperm protein (MSP), a small and abundant protein used in diagnostic immunoassays, which has been shown to be variable in some nematodes but not others. MSP was sequenced using worm DNA from eight adult worms from each of nine populations whose genetic structure previously had been quantified. For comparison, we also analyzed MSP sequences of the closely related Dictyocaulus eckerti and Dictyocaulus capreolus and from nematodes with sequences deposited in GenBank. In contrast to previous results, this study shows that the MSP of D. viviparus is similar to that of other nematodes. Almost no sequence variation, and thus no antigenic diversity, was detected in MSP between worms from different sub-populations or in the other Dictyocaulus species investigated. A functional test of a recombinant variant of the MSP showed that the expressed protein was recognized by antibodies in sera from infected cattle. This has practical implications for the development of species-specific markers, recombinant vaccines, and immunodiagnostics.

Molecular and Biochemical Parasitology, 1999

Numerous cathepsin B-like protein sequences (CBLs) have been reported from nematodes. However, the relationships among these proteins remain unclear. Here, expression of several CBL transcripts in the gut of the parasitic nematode Haemonchus contortus was demonstrated. To assess potential functional diversity, multiple nematode CBL sequences were compared with known functional domains of cathepsin B. These domains included the occluding loop, S2% and S2 subsites, and the pro region. Four groups of CBLs were defined based on variable characteristics in the occluding loop region, which incorporates a portion of the S2% subsite. Further diversity was observed in amino acids expected to contribute to the S2 subsite. In addition, short signature sequences near the cysteinyl active site region characterized known CBLs of parasites from the orders Strongylida and Rhabditida. The criteria established were used to identify two predicted CBLs from parasitic (Ascaris suum) and free-living (Caenorhabditis elegans) nematodes as potential orthologues, and provided a basis to evaluate orthologue status of other CBLs. Variability in the domains analyzed suggests substantial functional diversity in enzymatic properties of nematode CBLs. Results suggest that the selective amplification and evolution of distinct CBL lineages has contributed to differences in CBLs among species and groups of nematodes. Nutrient digestion is one potential factor promoting CBL diversification in these organisms.

Molecular and Biochemical Parasitology, 1999

Invading infective third-stage larvae (L 3 ) of parasitic nematodes execute a series of programmed developmental events in response to a host-specific signal encountered during infection. One of these early events is the release of excretory/secretory products. Using an in vitro feeding assay that mimics these early events of infection, a protein released by in vitro activated larvae of the hookworm Ancylostoma caninum was identified. This protein, Ac-ASP-2, was partially sequenced, and the cDNA encoding it isolated by PCR and screening of an A. caninum L 3 cDNA library. The Ac-asp-2 cDNA encodes a protein of 219 amino acids that is related to a previously identified protein, Ac-ASP-1, from hookworms. Both molecules are members of an evolutionarily diverse family of molecules that include the venom allergens of the Hymenoptera, and the testes specific proteins/sperm-coating glycoproteins of mammals. Homologues are present in nearly all nematodes tested, as demonstrated by PCR-hybridization and database searching. The Ac-asp-2 mRNA is synthesized in all life history stages, but the gene product is released only by L 3 activated to feed in vitro. The wide distribution of the Ac-asp-2 in nematodes and its release in response to host specific signals suggests that Ac-ASP-2 serves an important function in nematode physiology and development, and possibly in the infective process of parasitic species. (J.M. Hawdon) 0166-6851/99/$ -see front matter © 1999 Elsevier Science B.V. All rights reserved. PII: S 0 1 6 6 -6 8 5 1 ( 9 9 ) 0 0 0 1 1 -0