: Our research is aimed at understanding gene expression in the family Trypanosomatidae, which includes the parasitic protozoon's responsible for leishmaniasis, African Sleeping Sickness, and Chagas Disease. The focus is on the genesis and function of the spliced leader (SL) RNA, a small RNA that contributes the 5'-end sequence to every nuclear messenger RNA via a trans-splicing reaction. Trans-splicing, which is necessary for the conversion of polycistronic pre-messenger RNA (a likely consequence of the unusual genome organization found in kinetoplastids) into monocistronic mRNA, is not found in the human host or insect vector and thus represents a possible therapeutic target. This proposal outlines three complementary sets of experiments that detail the maturation pathway of the SL RNA common to Leishmania tarentolae and Trypanosoma brucei. 1) To refine the map of cis elements (SL RNA sequences and structures) necessary for accurate and efficient maturation and trans-splicing. Data obtained will allow the future identification of trans factors (protein and RNA) that interact with essential regions of RNA. 2) To isolate and examine the role of an XPO1-dependent nuclear export complex in SL RNA trafficking. 3) Experimental examination of the roles of a 3'-exonuclease and two ribose 2'-O- methyltransferases in the maturation of the 3' and 5' ends of the SL RNA, and bioinformatic identification and modeling of these proteins' interaction with the SL RNA. These three goals address discrete steps in the maturation of the SL RNA that determine its subsequent function. Specific inhibition of the trans-splicing pathway is lethal to the cell and thus a prime target for anti-parasite intervention.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI056034-04
Application #
7196546
Study Section
Tropical Medicine and Parasitology Study Section (TMP)
Program Officer
Mcgugan, Glen C
Project Start
2004-04-01
Project End
2009-03-31
Budget Start
2007-04-01
Budget End
2008-03-31
Support Year
4
Fiscal Year
2007
Total Cost
$353,390
Indirect Cost
Name
University of California Los Angeles
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Freire, Eden R; Sturm, Nancy R; Campbell, David A et al. (2017) The Role of Cytoplasmic mRNA Cap-Binding Protein Complexes in Trypanosoma brucei and Other Trypanosomatids. Pathogens 6:
Lin, Senjie; Cheng, Shifeng; Song, Bo et al. (2015) The Symbiodinium kawagutii genome illuminates dinoflagellate gene expression and coral symbiosis. Science 350:691-4
Yang, Feifei; Xu, Donghui; Zhuang, Yunyun et al. (2015) Spliced leader RNA trans-splicing discovered in copepods. Sci Rep 5:17411
Freire, Eden R; Malvezzi, Amaranta M; Vashisht, Ajay A et al. (2014) Trypanosoma brucei translation initiation factor homolog EIF4E6 forms a tripartite cytosolic complex with EIF4G5 and a capping enzyme homolog. Eukaryot Cell 13:896-908
Freire, Eden R; Vashisht, Ajay A; Malvezzi, Amaranta M et al. (2014) eIF4F-like complexes formed by cap-binding homolog TbEIF4E5 with TbEIF4G1 or TbEIF4G2 are implicated in post-transcriptional regulation in Trypanosoma brucei. RNA 20:1272-86
Porcel, Betina M; Denoeud, France; Opperdoes, Fred et al. (2014) The streamlined genome of Phytomonas spp. relative to human pathogenic kinetoplastids reveals a parasite tailored for plants. PLoS Genet 10:e1004007
Zhang, Huan; Campbell, David A; Sturm, Nancy R et al. (2013) Signal recognition particle RNA in dinoflagellates and the Perkinsid Perkinsus marinus. Protist 164:748-61
Zingales, Bianca; Miles, Michael A; Campbell, David A et al. (2012) The revised Trypanosoma cruzi subspecific nomenclature: rationale, epidemiological relevance and research applications. Infect Genet Evol 12:240-53
Tuller, Tamir; Girshovich, Yana; Sella, Yael et al. (2011) Association between translation efficiency and horizontal gene transfer within microbial communities. Nucleic Acids Res 39:4743-55
Werner, Maria; Purta, Elzbieta; Kaminska, Katarzyna H et al. (2011) 2'-O-ribose methylation of cap2 in human: function and evolution in a horizontally mobile family. Nucleic Acids Res 39:4756-68

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