Gal4-p300

Relevance: p300 is a transcriptional coactivator that functions as an integrator of numerous signaling pathways and are utilized by many DNA binding proteins to facilitate transcriptional activation. p300 shares numerous conserved domains with CREB binding protein (CBP), which is also a transcriptional coactivator. These shared domains include a histone acetyl transferase (HAT) domain, a bromo domain, and three cysteine- and histidine-rich domains. CBP also interacts with the RNA polymerase II holoenzyme and p300/CBP both contain transcriptional activation domains that function independently of HAT activity.
Description:

Gal4-p300

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Recommended Growth Conditions:

HeLa, U-2 OS and COS7 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% Fetal calf serum (FCS). HeLa His6-SUMO-1 stable cell line was grown in DMEM supplemented with 10% FCS and puromycin (2 g/ml).

References:

Girdwood D et al. 2003. Mol Cell. 11:1043-54. PMID: 12718889 Read more

Notes:

Calcium phosphate-mediated transfection of U-2OScells was performed with Gal4 p300 fusion proteins and subsequently verified with CAT or luciferase assays in U-2 OS cells. A total of 12g of plasmid DNA encoding one of HA-SUMO-1, HA-SUMO-2, or HA-SUMO-3 with either His6-p300(FL) or His6-p300(CRD)were cotransfected using Lipofectamine into COS7 cells.