Functional interaction between coactivators CBP/p300, PCAF, and transcription factor FKLF2

Area of Science:

• Molecular Biology
• Gene Regulation
• Transcription Factors

Background:

• The Sp1/KLF family regulates crucial cellular processes.
• Fetal Krüppel-like factor 2 (FKLF2) is a recently identified member of this family.
• Understanding FKLF2's transcriptional regulation is key to deciphering its role in growth and development.

Purpose of the Study:

• To identify and characterize coactivators involved in FKLF2 transcriptional activation.
• To elucidate the specific roles of CBP/p300 and PCAF in FKLF2 function.
• To investigate the mechanism by which coactivators enhance FKLF2 activity.

Main Methods:

• Co-transfection assays to measure transcriptional activity.
• Western blotting and co-immunoprecipitation to assess protein interactions.
• In vitro acetylation assays to determine enzyme activity.
• Electrophoretic mobility shift assays (EMSAs) to evaluate DNA binding.

Main Results:

• Both CBP/p300 and PCAF significantly enhance FKLF2 transcriptional activity.
• PCAF's acetyltransferase activity is essential for FKLF2 transcription, while CBP/p300's is not.
• CBP/p300 and PCAF cooperate to boost FKLF2 transcriptional activation.
• FKLF2 interacts with CBP and PCAF, and is acetylated by both.
• Both coactivators enhance FKLF2 DNA binding, with PCAF's acetyltransferase domain being critical.

Conclusions:

• CBP/p300 and PCAF are key coactivators for FKLF2.
• Distinct roles of acetyltransferase activities of CBP/p300 and PCAF in FKLF2 transcription and DNA binding.
• Enhanced DNA binding is a primary mechanism for FKLF2 transcriptional stimulation by these coactivators.