The proposed research focuses on the RNA capping reaction in trypanosomes. The investigator has purified the capping enzyme (guanylyltransferase) from Crithidia fasciculata and has cloned the corresponding genes from C. fasciculata and Trypanosoma brucei. The long-term goal of the proposed research is to understand the mechanism and the basis for the RNA specificity of the capping reaction of trypanosomatids. The investigator proposes three specific aims: (1) to undertake a structure-function analysis of the trypanosome capping enzyme; (2) to assess the consequences of genetic depletion of the capping enzyme; and (3) to search for cellular components that physically interact with the capping enzyme.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI043594-03
Application #
6170978
Study Section
Tropical Medicine and Parasitology Study Section (TMP)
Program Officer
Fairfield, Alexandra
Project Start
1998-06-01
Project End
2002-05-31
Budget Start
2000-06-01
Budget End
2001-05-31
Support Year
3
Fiscal Year
2000
Total Cost
$303,794
Indirect Cost
Name
Yale University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Kolev, Nikolay G; Ullu, Elisabetta; Tschudi, Christian (2015) Construction of Trypanosoma brucei Illumina RNA-Seq libraries enriched for transcript ends. Methods Mol Biol 1201:165-75
Ramey-Butler, Kiantra; Ullu, Elisabetta; Kolev, Nikolay G et al. (2015) Synchronous expression of individual metacyclic variant surface glycoprotein genes in Trypanosoma brucei. Mol Biochem Parasitol 200:1-4
Ericson, Megan; Janes, Michael A; Butter, Falk et al. (2014) On the extent and role of the small proteome in the parasitic eukaryote Trypanosoma brucei. BMC Biol 12:14
Kolev, Nikolay G; Ullu, Elisabetta; Tschudi, Christian (2014) The emerging role of RNA-binding proteins in the life cycle of Trypanosoma brucei. Cell Microbiol 16:482-9
Shi, Huafang; Barnes, Rebecca L; Carriero, Nicholas et al. (2014) Role of the Trypanosoma brucei HEN1 family methyltransferase in small interfering RNA modification. Eukaryot Cell 13:77-86
Atayde, Vanessa D; Ullu, Elisabetta; Kolev, Nikolay G (2012) A single-cloning-step procedure for the generation of RNAi plasmids producing long stem-loop RNA. Mol Biochem Parasitol 184:55-8
Kolev, Nikolay G; Ramey-Butler, Kiantra; Cross, George A M et al. (2012) Developmental progression to infectivity in Trypanosoma brucei triggered by an RNA-binding protein. Science 338:1352-3
Kolev, Nikolay G; Franklin, Joseph B; Carmi, Shai et al. (2010) The transcriptome of the human pathogen Trypanosoma brucei at single-nucleotide resolution. PLoS Pathog 6:e1001090
Patrick, Kristin L; Shi, Huafang; Kolev, Nikolay G et al. (2009) Distinct and overlapping roles for two Dicer-like proteins in the RNA interference pathways of the ancient eukaryote Trypanosoma brucei. Proc Natl Acad Sci U S A 106:17933-8
Shi, Huafang; Chamond, Nathalie; Djikeng, Appolinaire et al. (2009) RNA interference in Trypanosoma brucei: role of the n-terminal RGG domain and the polyribosome association of argonaute. J Biol Chem 284:36511-20

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