Identification In Vivo of Different Rate-Limiting Steps Associated with Transcriptional Activators in the Presence and Absence of a GAGA Element

@article{citeulike:2958782, abstract = {We analyzed the impact of a GAGA element on a transgenic promoter in Drosophila melanogaster that was activated by proteins composed of the Teton DNA binding domain and either the heat shock factor (HSF) activation domain or a potent subdomain of VP16. Permanganate footprinting was used to monitor polymerase II (Pol II) on the transgenic promoters in vivo. Activation by Teton-HSF but not by Teton-VP16A2 required the GAGA element; this correlated with the ability of the GAGA element to establish a paused Pol II. Although the GAGA element was not required for activation by Teton-VP16A2, the GAGA element greatly accelerated the rate of activation. The permanganate data also provided evidence that Pol II encountered different rate-limiting steps, following initiation in the presence of Teton-HSF and Teton-VP16A2. The rate-limiting step in the presence of Teton-HSF was release of Pol II paused about 20 to 40 nucleotides downstream from the start site. The rate-limiting step in the presence of Teton-VP16A2 occurred much closer to the transcription start site. Several biochemical studies have provided evidence for a structural transition shortly after Pol II initiates transcription. The behavior of Pol II in the presence of Teton-VP16A2 provides the first evidence that this transition occurs in vivo. 10.1128/MCB.25.9.3543-3552.2005}, author = {Wang, Yunyuan V. and Tang, Hongbing and Gilmour, David S. }, citeulike-article-id = {2958782}, doi = {http://dx.doi.org/10.1128/MCB.25.9.3543-3552.2005}, journal = {Mol. Cell. Biol.}, keywords = {initiation, pause, promoter-proximal, transcription}, month = {May}, number = {9}, pages = {3543--3552}, posted-at = {2008-07-03 12:14:14}, priority = {2}, title = {Identification In Vivo of Different Rate-Limiting Steps Associated with Transcriptional Activators in the Presence and Absence of a GAGA Element}, url = {http://dx.doi.org/10.1128/MCB.25.9.3543-3552.2005}, volume = {25}, year = {2005} }

TY - JOUR ID - citeulike:2958782 L3 - citeulike-article-id:2958782 N2 - We analyzed the impact of a GAGA element on a transgenic promoter in Drosophila melanogaster that was activated by proteins composed of the Teton DNA binding domain and either the heat shock factor (HSF) activation domain or a potent subdomain of VP16. Permanganate footprinting was used to monitor polymerase II (Pol II) on the transgenic promoters in vivo. Activation by Teton-HSF but not by Teton-VP16A2 required the GAGA element; this correlated with the ability of the GAGA element to establish a paused Pol II. Although the GAGA element was not required for activation by Teton-VP16A2, the GAGA element greatly accelerated the rate of activation. The permanganate data also provided evidence that Pol II encountered different rate-limiting steps, following initiation in the presence of Teton-HSF and Teton-VP16A2. The rate-limiting step in the presence of Teton-HSF was release of Pol II paused about 20 to 40 nucleotides downstream from the start site. The rate-limiting step in the presence of Teton-VP16A2 occurred much closer to the transcription start site. Several biochemical studies have provided evidence for a structural transition shortly after Pol II initiates transcription. The behavior of Pol II in the presence of Teton-VP16A2 provides the first evidence that this transition occurs in vivo. 10.1128/MCB.25.9.3543-3552.2005 IS - 9 JF - Mol. Cell. Biol. EP - 3552 TI - Identification In Vivo of Different Rate-Limiting Steps Associated with Transcriptional Activators in the Presence and Absence of a GAGA Element VL - 25 SP - 3543 KW - initiation KW - pause KW - promoter-proximal KW - transcription AU - Wang, Yunyuan AU - Tang, Hongbing AU - Gilmour, David PY - 2005/05/01/ UR - http://dx.doi.org/10.1128/MCB.25.9.3543-3552.2005 ER -