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    Characterization of a Novel Class I Transcription Factor A (CITFA) Subunit That Is Indispensable for Transcription by the Multifunctional RNA Polymerase I of Trypanosoma brucei

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    Type
    Article
    Authors
    Nguyen, T. N.
    Nguyen, B. N.
    Lee, J. H.
    Panigrahi, Aswini Kumar
    Gunzl, A.
    KAUST Department
    Bioscience Core Lab
    Date
    2012-10-26
    Online Publication Date
    2012-10-26
    Print Publication Date
    2012-12
    Permanent link to this record
    http://hdl.handle.net/10754/554100
    
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    Abstract
    Trypanosoma brucei is the only organism known to have evolved a multifunctional RNA polymerase I (pol I) system that is used to express the parasite's ribosomal RNAs, as well as its major cell surface antigens, namely, the variant surface glycoprotein (VSG) and procyclin, which are vital for establishing successful infections in the mammalian host and the tsetse vector, respectively. Thus far, biochemical analyses of the T. brucei RNA pol I transcription machinery have elucidated the subunit structure of the enzyme and identified the class I transcription factor A (CITFA). CITFA binds to RNA pol I promoters, and its CITFA-2 subunit was shown to be absolutely essential for RNA pol I transcription in the parasite. Tandem affinity purification (TAP) of CITFA revealed the subunits CITFA-1 to -6, which are conserved only among kinetoplastid organisms, plus the dynein light chain DYNLL1. Here, by tagging CITFA-6 instead of CITFA-2, a complex was purified that contained all known CITFA subunits, as well as a novel proline-rich protein. Functional studies carried out in vivo and in vitro, as well as a colocalization study, unequivocally demonstrated that this protein is a bona fide CITFA subunit, essential for parasite viability and indispensable for RNA pol I transcription of ribosomal gene units and the active VSG expression site in the mammalian-infective life cycle stage of the parasite. Interestingly, CITFA-7 function appears to be species specific, because expression of an RNA interference (RNAi)-resistant CITFA-7 transgene from Trypanosoma cruzi could not rescue the lethal phenotype of silencing endogenous CITFA-7.
    Citation
    Characterization of a Novel Class I Transcription Factor A (CITFA) Subunit That Is Indispensable for Transcription by the Multifunctional RNA Polymerase I of Trypanosoma brucei 2012, 11 (12):1573 Eukaryotic Cell
    Publisher
    American Society for Microbiology
    Journal
    Eukaryotic Cell
    DOI
    10.1128/EC.00250-12
    PubMed ID
    23104567
    PubMed Central ID
    PMC3536272
    Additional Links
    http://ec.asm.org/cgi/doi/10.1128/EC.00250-12
    ae974a485f413a2113503eed53cd6c53
    10.1128/EC.00250-12
    Scopus Count
    Collections
    Articles; Bioscience Core Lab; Bioscience Core Lab

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