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    Cis-regulatory contributions to the regulation of sloppy-paired 1 transcription initiation and elongation

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    StonyBrookUniversityETDPageEmbargo_20130517082608_116839.pdf (40.31Kb)
    Date
    1-Aug-12
    Author
    Hang, Saiyu
    Publisher
    The Graduate School, Stony Brook University: Stony Brook, NY.
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    Abstract
    The expression of the sloppy-paired 1 (slp1) gene in the gastrula stage Drosophila embryo is controlled by the interplay of four transcription factors Runt, Even-skipped (Eve), Fushi-tarazu (Ftz), and Odd-paired (Opa) with two distinct cis-regulatory enhancer elements, the distal early stripe element (DESE) and the proximal early stripe element (PESE). The stripe pattern of slp1 is the result of non-additive interactions between these two enhancers and is context-dependent. Using chromatin immunoprecipitation (ChIP) to examine a number of reporter constructs, I found DESE mediates Runt dependent activation by facilitating pre-initiation complex formation on the slp1 promoter. This DESE-dependent activation is influenced by the extent of promoter- proximal DNA upstream of the transcription start site and involves a mechanism that induces nucleosome depletion around the promoter. This effect is specifically important for DESE activation but not PESE activation. ChIP experiments comparing wild-type versus repressed states of DESE-lacZ and PESE-lacZ reporter genes indicate that Eve represses PESE-lacZ expression by blocking the elongation step of the transcription cycle. This repression involves the regulated association of the elongation factor P-TEFb and phosphorylation of Ser2 in the C-terminal domain of RNA polymerase II. Runt and Ftz repress both DESE and PESE. Interestingly, Runt and Ftz repress DESE-lacZ by the same mechanism as Eve dependent repression of PESE, that is by inhibition of transcription elongation. However, Runt and Ftz repress PESE-lacZ by blocking transcription initiation. Analysis of the conserved domains of Runt revealed that C-terminal domain of Runt (region VIII) is involved in the repression of both DESE and PESE, and that this region is not required for activation of DESE.
    Description
    97 pg.
    URI
    http://hdl.handle.net/1951/60292
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    • Stony Brook Theses & Dissertations [SBU] [1956]

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