Category Archives: human pathogen

Recent animal-associated fungal genome papers

The genomes of five dermatophyte fungi were sequenced and the analyses of their lifestyles presented in a new paper out in mBio in Martinez et al. 2012. The authors were able to identify gene family changes that associate with lifestyle changes including proteases that can degrade keratin suggesting how these species have adapted to obtaining nutrients from an animal host. The continued finding of fungal-specific kinase families in these fungi, extending the observations from previous studies in Coprinopsis and Paracoccidioides on the FunK1 kinase family, makes me hope we will some day get some molecular information on the specificity of these families in addition to these copy number observations.
Another paper published in Genome Research this summer from Emily Troemel‘s lab and the Broad Institute describes the sequencing of two microsporidia species that are natural parasites of Caenorhabditis.The paper reveals some suprising things about Microsporidia evolution including the presence of a clade-specific nucleoside H+ symporter which is only found in bacteria and some eukaryotes and not in any Fungi. The phyletic distribution suggested it was acquired more recently and couple from lateral gene transfer. This acquisition likely helps the microsporidia cells obtain nucleosides from the host since the parasite cannot synthesize these. There is also evidence of evolution of microsporidia-specific secretion signals in the hexokinases which may be a mechanism for delivery of these enzymes into host cells to catalyze rapid growth once inside the host. Many more gems in this paper including phylogenetic placement of the microsporidia from phylogenomic approaches (also see related recent work from Toni Gabaldon‘s lab).

Microsporidia genomes on the way

New genomes from Microsporidia are on the way from the Broad Institute and other groups, and will be a boon to those working on these fascinating creatures. Microsporidia are obligate intracellular parasites of eukaryotic cells and many can cause serious disease in humans. Some parasitize worms and insects too. The evolutionary placement of these species in the fungi is still debated with recent evidence placing them as derived members of the Mucormycotina based on shared synteny (conserved gene order), in particular around the mating type locus.  There is still some debate as to where this group belongs in the Fungal kingdom, with their highly derived characteristics and long branches they are still make them hard to place.  The synteny-based evidence was another way to find a phylogenetic placement for them but it would be helpful to have additional support in the form of additional shared derived characteristics that group Mucormycotina and Microsporidia. There is hope that increased number of genome sequences and phylogenomic approaches can help resolve the placement and more further understand the evolution of the group.

For data analysis, a new genome database for comparing these genomes is online called MicrosporidiaDB. This project has begun incorporating the available genomes and providing a data mining interface that extends from the EuPathDB project.

Origins and evolution of pathogens An article in PLoS Pathogens by Morris et al describe a hypothesis about the evolution and origins of plant pathogens applying the parallel theories to the emergence of medically relevant pathogens. The authors highlight the importance of understanding the evolution of organisms in the context of emerging pathogens like Puccinia Ug99 for our ability to design strategies to protect human health and food supplies.  Both bacterial and fungal pathogens of plants are discussed but I (perhaps unsurprisingly) focus on the fungi here. Continue reading Origins and evolution of pathogens

Updated Cryptococcus serotype A annotation

SEM of clamp cell, yeast cells and sexual spore chains. Courtesy R. Velagapudi & J. Heitman

A new and improved annotation of Cryptococcus neoformans var grubii strain H99 (serotype A) has been made available in GenBank and the Broad Institute website. This update is collaboration between several groups providing data and analyses and the genome annotation team at the Broad Institute.

Some changes noted by the Broad Institute include:

“This release of gene predictions for the serotype A isolate Cryptococcus neoformans var. grubii H99 is based on a new genomic assembly provided by Dr. Fred Dietrich at the Duke Center for Genome Technology. The new assembly consists of 14 nuclear chromosomes and a single 21 KB mitochondrial chromosome, and has resulted in a reduction of the estimated genome size from 19.5 to 18.9 Mb. Improvements in the assembly and in our annotation process have resulted in a set of 6,967 predicted protein products, 335 fewer than the previous release.”

How to get A. fumigatus in the mood for love A manuscript at Nature AOP details the success of the Dyer lab and collaborators in encouraging Aspergillus fumigatus to complete the sexual cycle under observable (e.g. laboratory) conditions. The authors are the teleomorph (sexual or perfect) stage Neosartorya fumigata for a fungus that had been previously only had an observed anamorphic stage. A. fumigatus can reproduce asexually forming structures called conidiophores which produce asexual spores called conidiospores (or mitospores as they are produced via mitosis) define the anamorph or imperfect stage, but no sexual structures such as cleistothecia that produce the packaged sexual products as ascospores. See a presentation by David Geiser (archived at the Aspergillus website) for more detail on some of the morphological and phylogenetic characters that unify the group of Eurotiales fungi.

Like several other groups of fungi, A. fumigatus was presumed to have a putative cryptic sexual stages inferred from population genetic evidence of sexual recombination, but until no telemorphs had been observed. In addition, an observed perfect stage doesn’t necessarily indicate it is easy to induce mating in laboratory conditions. Complicated media including the ever stressful V8 juice was needed to induce mating in the basidiomycete yeast Cryptococcus neoformans (Erke, J Bacteriol 1976). In fact, Christina Hull’s lab has shown we still don’t even know what ingredients in V8 juice even induce mating (Kent et al, AEM 2008)! Other fungi including Coccidioides have been implicated as cryptically sexual (Burt et al, PNAS 1996) but no one has been able to induce mating in laboratory conditions. In this case a petri plate with a individual of each mating type (since this is a heterothallic fungus), and a series of different media conditions provided an environment suitable for mating to occur.

The work in this paper follows from their previous work identifying isolates of different mating types (Paoletti, Current Biol, 2005). The discovery of sexual stage for Aspergillus fumigatus (which Bret cannot pronounce) is a boon for molecular geneticists in construction of knockout strains and ability to follow recombination. While A. nidulans is a sexual species and model system for genetics, it is useful to have more tools to directly manipulate A. fumigatus and directly test hypotheses about genes involved in pathogenicity.

This observation of meiosis in the laboratory is also is interesting to considered in light of work that RIP is active in other Aspergillus species (and also see this post) suggesting that RIP may be operating under meiotic conditions.

Isolates of different mating types have also been described for the putatively asexual Coccidioiodes (Mandell et al, EC 2007; Fraser et al, EC 2007) so it remains a possibility that we can also induce sexual recombination in laboratory conditions in this fungus.

Céline M. O’Gorman, Hubert T. Fuller, Paul S. Dyer (2008). Discovery of a sexual cycle in the opportunistic fungal pathogen Aspergillus fumigatus Nature DOI: 10.1038/nature07528

Coprinus on the heart?

Here’s a fungal infection you don’t hear much about. One of the fungi we work on, a model for mushroom development as it can be fruited in the lab is Coprinopsis cinerea (previously named Coprinus cinereus). C. cinerea is a saprobric coprophillic fungus so it is usually found on dung.  Although rare in human infections there are a few reports in immunocopromised patients.  Below is an abstract describing isolation of C. cinerea from an implanted heart valve from a pig. This definitely not its typical habitat and Coprinus growing in yeast form I’m sure I’ve really heard of either.  Would be great to see if the clinical strains are still sexually competent and/or are significantly different in other ways (growth rate, resistance to drugs and oxidative stress) from the wild or laboratory strains.

A 77-year-old female initially presented with symptomatic mitral valve stenosis involving a bioprosthesis that had been implanted 8 months earlier for myxomatous mitral valve disease and severe valvular regurgitation. The patient was taken for a second mitral valve replacement due to stenosis. Intraoperatively, the bioprosthetic mitral valve was noted to have an unusual clot-like mass on the atrial side. Initial fungal smears were positive for yeast stains, and pathology revealed extensive colonization by thick filamentous fungus with apparent true hyphae, pseudohyphae, and yeast forms. The fungus was identified as Hormographiella aspergillata, the asexual form of Coprinus cinereus, a common inky cap mushroom that grows in the lawn.

Continue reading Coprinus on the heart?

Cryptococcus species deliniation What delineates species boundaries in fungi? Much work has been done on biological and phylogenetic species concepts in fungi. Some concepts are reviewed in Taylor et al 2006 and in Taylor et al 2000, and applications can be seen in several pathogens such as Paraccocidiodies, Coccidioides, and the model filamentous (non-pathogenic) fungus Neurospora.

A paper in Fungal Genetics and Biology on species definitions in Cryptococcus neoformans from multi-locus sequencing seeks to provide additional treatment of the observed diversity. A large study of 117 Cryptococcus isolates were examined through multi-locus sequencing (6 loci) and identified two monophyletic lineages within C. neoformans varieties that correspond to var. neoformans and var. grubii. However within the C. gattii samples they identified four monophyletic groups consistent with deep divergences observed from whole genome trees for two strains of C. gattii, MLST, and AFLP studies. By first defining species, we can now test whether any of the species groups have different traits including prevalence in clinical settings and in nature.

BOVERS, M., HAGEN, F., KURAMAE, E., BOEKHOUT, T. (2007). Six monophyletic lineages identified within Cryptococcus neoformans and Cryptococcus gattii by multi-locus sequence typing. Fungal Genetics and Biology DOI: 10.1016/j.fgb.2007.12.004

Saccharomyces strain sequencing

Blogging on Peer-Reviewed ResearchWhile many strains of S. cerevisiae are being sequenced, a single strain, YJM789, isolated from the lung of an AIDS patient was sequenced a few years ago at Stanford and published this summer. The genome was described in a paper entitled “Genome sequencing and comparative analysis of Saccharomyces cerevisiae strain YJM789”.

Continue reading Saccharomyces strain sequencing

Candida White-Opaque switching

Blogging on Peer-Reviewed ResearchA paper in PLoS Biology from Sandy Johnson’s lab entitled “Interlocking Transcriptional Feedback Loops Control White-Opaque Switching in Candida albicans discusses phenotype switching in the human pathogenic fungus Candida albicans. Why is the important?

“White-opaque switching is an epigenetic phenomenon, where genetically identical cells can exist in two distinctive cell types, white and opaque. Each cell type is stably inherited for many generations, and switching between the two types of cells occurs stochastically and rarely—roughly one switch in 10^4 cell divisions”

white-opaque coloniesThere is also a review by Kira O’Day to discuss the implications of the findings. Understanding this sort of developmental and epigenetic signaling is important to better know how fungi adjust and interact with their environment. However, the authors do conclude that White-Opaque switching is exclusive to Candida albicans so aspects of this research only directly applicable to studies in this system. Phenotype switching is an active area of research for Candida biologists – some nice micrographs and SEM of the different cell morphologies can be seen at Prof. Joachim Morschhäuser’s page (and linked to the right).

Continue reading Candida White-Opaque switching

Evolution of aflatoxin gene cluster

Blogging on Peer-Reviewed ResearchIgnazio Carbone and colleagues published a recent analysis of the evolution of the aflatoxin gene cluster in five Aspergillus fungi entitled “Gene duplication, modularity and adaptation in the evolution of the aflatoxin gene cluster” in BMC Evolutionary Biology. The authors were able to identify seven modules pairs of genes whose history of duplication were highly correlated. Several genomes of Aspergillus have been sequenced along with more Eurotioales fungi. Continue reading Evolution of aflatoxin gene cluster