Toxoplasma Gondii Developmental Biology: A Correlative Transcriptome Proteome Approach
EMSL Project ID
40070
Abstract
The Apicomplexa include most of the major protozoan pathogens of humans and animals, such as Plasmodia (malaria), Babesia, Theileria, Cryptosporidia, Cyclospora, Neospora, Eimeria, and Isospora. Of these organisms, Toxoplasma gondii is a ubiquitous parasite of warm-blooded animals and is one of the most common parasitic infections of humans. It has been estimated that 1 per 1000 children born each year in the United States have congenital toxoplasmosis. T. gondii is also recognized as a pathogen of immune compromised hosts. Due to the ease with which T. gondii can be manipulated within the laboratory and the availability of techniques for genetic transformation, T. gondii can be used to study many aspects of Apicomplexan biology that cannot be readily studied in these other organisms. A major focus in the study of intracellular pathogens is the changes that occur in transcription and protein expression when these organisms undergo differentiation or respond to environmental signals (including host immune factors). Differentiation is a key process in the success of the Apicomplexa, with these organisms having separate life cycle stages associated with transmission of infection as well as tissue residence. The molecular mechanisms underlying this process are not clearly understood. A powerful approach for understanding the differentiation process is to use mRNA and protein expression profiling. The relationship between mRNA and protein expression is, however, poorly understood in the Apicomplexa, as well as other organisms. The ability to relate quantitative changes in the proteome with quantitative changes in the transcriptome has the potential to dramatically enhance our understanding of gene regulation and the events of differentiation in important pathogen systems. In addition, studies of the vacuole membrane in which parasites reside can provide invaluable data on the interface between intracellular pathogens and their host cells. A major limitation of these studies, in the past, has been the small amount of material available for transcriptional or proteomic analyses. With the development of newer methods such as RNA-Seq and highly sensitive mass spectrometry instruments, the problem of small sample size is less of an issue. Thus, important host-pathogen issues can be addressed in a systematic fashion and at the genome scale. We propose to do a detailed correlative transcriptome, using RNA seq, and quantitative proteome, using mass spectrometry, of the developmental biology of T. gondii providing a dynamic profile of differentiation in this pathogen. The development and refinement of the experimental techniques used in this proposal will also be applicable to many systems in which limitation of mass is a problem or when intracellular residence of the organism of interest is an issue.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2010-10-01
End Date
2013-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Croken M, YF Ma, LM Markillie, RC Taylor, G Orr, LM Weiss, and K Kim. 2014. "Distinct Strains of Toxoplasma gondii Feature Divergent Transcriptomes Regardless of Developmental Stage." PLoS One 9(11):, doi:10.1371/journal.pone.0111297
Suvorova ES, M Croken, S Kratzer, LM Ting, M Conde de Felipe, B Balu, LM Markillie, LM Weiss, K Kim, and MW White. 2013. "Discovery of a Splicing Regulator Required for Cell Cycle Progression." PLoS Genetics 9(2):Article No. e1003305. doi:10.1371/journal.pgen.1003305
Tomita T, DJ Bzik, YF Ma, BA Fox, LM Markillie, RC Taylor, K Kim, and LM Weiss. 2013. "The Toxoplasma gondii cyst wall protein CST1 is critical for cyst wall integrity and promotes bradyzoite persistence." PLoS Pathogens 9(12):e1003823. doi:10.1371/journal.ppat.1003823