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Nuclear proteins from Leishmania tarentolae can bind to a putative tRNA promoter element
Author(s) -
Jennings Kimberly,
Alfonzo Juan,
Pitula Joseph
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1003.14
Subject(s) - transfer rna , oligonucleotide , biology , rna polymerase iii , transcription (linguistics) , leishmania , rna , gene , dna , promoter , leishmania major , microbiology and biotechnology , genetics , rna polymerase , gene expression , linguistics , philosophy , parasite hosting , world wide web , computer science
Trypanosomatids are the cause of many diseases across the world. Leishmania is a specific trypanosomatid that is the causative agent of Leishmaniasis. Trypanosomes are unique in their tRNA transcription in that, two transcription factors, TFIIIC and TFIIIB, which were believed to be necessary for RNA Pol III recognition, are not found in genomic databases. To find out if there are any tRNA transcription factors that bind to the promoter region of the tRNA gene, we conducted band‐shift assays using nuclear extracts from Leishmania cultures and an oligonucleotide sequence of the promoter region. The assays revealed three distinct protein‐ DNA complexes. By using increasing amounts of NaCl, we found that higher levels of salt (>0.15M) lead to an inhibition of protein binding. We also used competition assays to test the specificity of the complexes using unlabeled duplex and a PCR fragment. We conclude that complex C is the most stable complex and may not be specific to the oligonucleotide sequence. Further assays will need to be conducted to characterize the two remaining complexes.