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Dissertation
Characterization of PNUDC: a plasmodial homolog of a protein essential for nuclear migration in the fungus Aspergillus nidulans
Doctor of Philosophy (Ph.D.), Medical College of Pennsylvania and Hahnemann University
Sep 2000
DOI:
https://doi.org/10.17918/00007644
Abstract
In recent years, malaria has reemerged as a deadly global disease. No working vaccine against Plasmodium, the parasite that causes malaria, is available. Furthermore, parasite resistance to currently available drugs is rapidly emerging. A limited knowledge of the intricacies of Plasmodium's cell biology contributes to our inability to develop better chemotherapeutics. Nuclear migration is important in many developmental processes and cell cycle events, In Plasmodium, nuclear migration is paramount to propagation of its complex life cycle, and varies from that in the vertebrate host. At least two distinct times in the life cycle (schizogony and sporogony), nuclei must migrate in an orderly, concerted fashion. Understanding the mechanisms responsible for these processes could lead to new plasmodial drug targets. NudC is a protein essential for nuclear migration in Aspergillus nidulans. Homologs of NudC are reported in Drosophila, mouse, rat, humans, C. elegans, and S. pombe, attesting to its conserved role throughout evolution. All homologs share remarkable sequence homology at the carboxy terminus. Using a combination of PCR techniques and computer homolog alignment searches, P. yoelii nudC (pynudC) and P. falciparum nudC (pfnudC) were identified and sequenced. The genes were characterized via Southern and Northern analyses, and RT-PCR. Computer analysis was conducted on the proteins, including 1° and 2° structure motif searches, and alignments. PynudC expression for complementation, the development of antibodies, and protein-protein interaction studies were conducted. Additionally, Western analysis, immunoprecipitations and immunofluorescence assays were performed on the erythrocytic stages of the parasite. Both pynudC and pfnudC are intronless single-copy genes. They are transcribed during the erythrocytic stages of the life cycle. PfnudC and PynudC proteins are present during the erythrocytic stages, with greatest expression seen during schizogony and in gametocytes. Expression is observed in the cytosol. The proteins are predicted to be hydrophilic, supporting a cytosolic location within the cell. They do not share identifiable motifs to other known protein families. These findings support the hypothesis that PnudC is necessary during nuclear migration in Plasmodium. They should lead to an improved understanding of the intracellular events leading to schizogony and sporogony, and may therefore contribute to novel drug developments.
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Details
- Title
- Characterization of PNUDC
- Creators
- Sharon Lewis Bloomgarden
- Contributors
- Carole A. Long (Advisor) - Drexel University, Medical College of Pennsylvania and Hahnemann University (1993-1996, 1998-2002)
- Awarding Institution
- Medical College of Pennsylvania and Hahnemann University
- Degree Awarded
- Doctor of Philosophy (Ph.D.)
- Publisher
- Medical College of Pennsylvania and Hahnemann University; Philadelphia, Pennsylvania
- Number of pages
- ix, 139 pages
- Resource Type
- Dissertation
- Language
- English
- Academic Unit
- Medical College of Pennsylvania and Hahnemann University (1993-1996, 1998-2002)
- Other Identifier
- 991021888810704721