We have shown that two trypanosome-specific RNA binding proteins, P34 and P37, play an essential role in the ribosomal biogenesis pathway in T. brucei. This represents a novel finding since the ribosomal pathway is highly conserved. To exploit this observation for the ultimate goal of targeting the T. brucei ribosomal pathway in chemotherapy, we must understand the mechanism by which these proteins act in the pathway. In recent work, we have shown that P34 and P37 interact with both L5 and 5S rRNA, the highly conserved components of the preribosomal particle. Loss of P34/P37 leads to a decrease in 5S rRNA levels and other effects in ribosomal formation. The objective of this proposal is to determine the critical residues involved in this interaction and develop a method to identify molecules that disrupt this interaction. Our hypothesis is that the molecular interactions of these proteins will provide a valid target for intervention in ribosomal assembly in T. brucei.
The specific aims of the project are to: 1. Determine the critical molecular determinants on L5 and P34/P37 that are required for participation in the T. brucei preribosomal complex. 2. Determine the critical regions on 5S rRNA that interact with L5 and P34/P37 in the T. brucei preribosomal complex. 3. Determine whether the interaction of P34/P37 with L5 and 5S rRNA is required for trafficking of the complex to the nucleolus. 4. Develop a FRET (fluorescence energy transfer) based screen for the disruption of the interaction between P34/P37, L5, and 5S rRNA in the preribosomal particle. Differences in ribosomal structure and function between bacteria and the infected human host have provided targets for macrolide and ketolide antibiotics. Since this preribosomal particle is unique to kinetoplastids and play an essential role in the ribosomal structure and biogenesis in these organisms, mechanistic studies with this complex will allow us to define a strategic target for chemical intervention.

Public Health Relevance

The protozoan parasites, the Kinetoplastids, include the organisms Trypanosoma brucei, Trypanosoma cruzi and Leishmania, which cause serious diseases in much of the world (African sleeping sickness, Chagas'disease and leishmaniasis, respectively). This study will examine a unique preribosomal complex occurring in these parasites with the goal of developing an assay to identify drugs that target the parasite but not host complex.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM092719-03
Application #
8247798
Study Section
Pathogenic Eukaryotes Study Section (PTHE)
Program Officer
Bender, Michael T
Project Start
2010-04-01
Project End
2014-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
3
Fiscal Year
2012
Total Cost
$309,435
Indirect Cost
$111,435
Name
State University of New York at Buffalo
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
038633251
City
Buffalo
State
NY
Country
United States
Zip Code
14260
Umaer, Khan; Ciganda, Martin; Williams, Noreen (2014) Ribosome biogenesis in african trypanosomes requires conserved and trypanosome-specific factors. Eukaryot Cell 13:727-37
Wang, Lei; Ciganda, Martin; Williams, Noreen (2013) Defining the RNA-protein interactions in the trypanosome preribosomal complex. Eukaryot Cell 12:559-66
Wang, Lei; Ciganda, Martin; Williams, Noreen (2013) Association of a novel preribosomal complex in Trypanosoma brucei determined by fluorescence resonance energy transfer. Eukaryot Cell 12:322-9
Sakyiama, Joseph; Zimmer, Sara L; Ciganda, Martin et al. (2013) Ribosome biogenesis requires a highly diverged XRN family 5'->3' exoribonuclease for rRNA processing in Trypanosoma brucei. RNA 19:1419-31
Ciganda, Martin; Williams, Noreen (2012) Characterization of a novel association between two trypanosome-specific proteins and 5S rRNA. PLoS One 7:e30029
Ciganda, Martin; Williams, Noreen (2011) Eukaryotic 5S rRNA biogenesis. Wiley Interdiscip Rev RNA 2:523-33