We have sequenced the genome of the emerging human pathogen
Babesia microti
and compared it with that of other protozoa.
B. microti
has the smallest nuclear genome among all Apicomplexan parasites sequenced to date with three chromosomes encoding ∼3500 polypeptides, several of which are species specific. Genome-wide phylogenetic analyses indicate that
B. microti
is significantly distant from all species of Babesidae and Theileridae and defines a new clade in the phylum Apicomplexa. Furthermore, unlike all other Apicomplexa, its mitochondrial genome is circular. Genome-scale reconstruction of functional networks revealed that
B. microti
has the minimal metabolic requirement for intraerythrocytic protozoan parasitism.
B. microti
multigene families differ from those of other protozoa in both the copy number and organization. Two lateral transfer events with significant metabolic implications occurred during the evolution of this parasite. The genomic sequencing of
B. microti
identified several targets suitable for the development of diagnostic assays and novel therapies for human babesiosis.
Sphingolipids are ubiquitous and essential components of eukaryotic membranes, particularly the plasma membrane. The biosynthetic pathway for the formation of these lipid species is conserved up to the formation of sphinganine. However, a divergence is apparent in the synthesis of complex sphingolipids. In animal cells, ceramide is a substrate for sphingomyelin (SM) production via the enzyme SM synthase. In contrast, fungi utilize phytoceramide in the synthesis of inositol phosphorylceramide (IPC) catalyzed by IPC synthase. Because of the absence of a mammalian equivalent, this essential enzyme represents an attractive target for anti-fungal compounds. In common with the fungi, the kinetoplastid protozoa (and higher plants) synthesize IPC rather than SM. However, orthologues of the gene believed to encode the fungal IPC synthase (AUR1) are not readily identified in the complete genome data bases of these species. By utilizing bioinformatic and functional genetic approaches, we have isolated a functional orthologue of AUR1 in the kinetoplastids, causative agents of a range of important human diseases. Expression of this gene in a mammalian cell line led to the synthesis of an IPC-like species, strongly indicating that IPC synthase activity is reconstituted. Furthermore, the gene product can be specifically inhibited by an anti-fungal-targeting IPC synthase. We propose that the kinetoplastid AUR1 functional orthologue encodes an enzyme that defines a new class of protozoan sphingolipid synthase. The identification and characterization of the protozoan IPC synthase, an enzyme with no mammalian equivalent, will raise the possibility of developing anti-protozoal drugs with minimal toxic side affects.
Graphical abstractHighlights► Identification and characterisation of Toxoplasma sphingolipid synthase (TgSLS). ► Demonstration of TgSLS inositol phosphorylceramide synthase activity. ► Identification of inositol phosphorylceramide in Toxoplasma extracts. ► Delineation of role of host sphingolipid biosynthesis in Toxoplasma proliferation. ► Host biosynthesis non-essential for proliferation, de novo synthesis could be key.
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AbstractSphingolipids are important components of eukaryotic membranes, particularly the plasma membrane, and are involved in a diverse array of signal transduction processes.In the Eukaryota the biosynthetic pathway for the formation of these lipid species is largely conserved. However, in contrast to mammals which produce sphingomyelin (SM), several pathogenic fungi and protozoa synthesize inositol phosphorylceramide 3
Key wordsTrypanosoma, trypanosomiasis, sphingolipid synthase, drug target
FootnoteSince the initial submission of this work to Molecular and Biochemical Parasitology a paper has been published in Molecular Microbiology identifying and characterising the same family of enzymes in Trypanosoma brucei:
The Mv1751 gene product is thought to catalyze the first step in the N-glycosylation pathway in Methanococcus voltae. Here, we show that a conditional lethal mutation in the alg7 gene (N-acetylglucosamine-1-phosphate transferase) in Saccharomyces cerevisiae was successfully complemented with Mv1751, highlighting a rare case of cross-domain complementation.
P.W. (2010) 'A plate-based assay system for analyses and screening of the Leishmania major inositol phosphorylceramide synthase.', International journal of biochemistry and cell biology., 42 (9). pp. 1553-1561. Further information on publisher's website:https://doi.org/10. 1016/j.biocel.2010.06.008 Publisher's copyright statement: NOTICE: this is the author's version of a work that was accepted for publication in International journal of biochemistry and cell biology.
Additional information:Use policyThe full-text may be used and/or reproduced, and given to third parties in any format or medium, without prior permission or charge, for personal research or study, educational, or not-for-prot purposes provided that:• a full bibliographic reference is made to the original source • a link is made to the metadata record in DRO • the full-text is not changed in any way The full-text must not be sold in any format or medium without the formal permission of the copyright holders.Please consult the full DRO policy for further details. N-[6-[(7-nitro-2-1,3-benzoxadiazol-4-yl) Furthermore, inhibitory substrate analogues have been identified. Importantly this assay is amenable to development for use in high-throughput screening applications for lead inhibitors and as such may prove to be a pivotal tool in drug discovery.3
Further information on publisher's website:http://dx.doi.org/10.1007/s11103-010-9626-3Publisher's copyright statement:The original publication is available at www.springerlink.com Additional information:
Use policyThe full-text may be used and/or reproduced, and given to third parties in any format or medium, without prior permission or charge, for personal research or study, educational, or not-for-prot purposes provided that:• a full bibliographic reference is made to the original source • a link is made to the metadata record in DRO • the full-text is not changed in any way The full-text must not be sold in any format or medium without the formal permission of the copyright holders.Please consult the full DRO policy for further details. Here we functionally analysed all three predicted Arabidopsis IPC synthases, confirming them as aureobasidin A resistant AUR1p orthologues. Expression profiling revealed that the genes encoding these orthologues are differentially expressed in various tissue types isolated from Arabidopsis.
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