| Popis: |
The function of transcription extension factor 1 (elf1) protein was mainly studied by Saccharomyces cerevisiae, but it was rarely reported in low eukaryotes. In addition, the role of Physarum ELF1 (PELF1) and PELF1/Physarum 14-3-3 (P14-3-3) protein interaction is poorly understood. Q-PCR analysis of the PELF1 mRNA levels indicates that it involved DNA structure formation. Further, confirmed that the DNA binding site of PELF1 is located within 1 to 22 amino acids from the N-terminus. The site of the interaction between PELF1 and P14-3-3 is located on the 40-RDAGI-44 aa peptide at the corner of the βA→βB of the C4 zinc finger domain. Key peptides and amino acids affecting PELF1 binds to DNA. Electrophoretic mobility shift analysis (EMSA) results showed that PELF is faster than P14-3-3/PELF1 complex when they binding to PardC. Atomic force microscopy (AFM) showed that both PELF1/DNA complex and P14-3-3/PELF1/DNA complex aggregated, but P14-3-3/PELF1/DNA complex aggregated most. ITC (Isothermal Titration Calorimetry) showed that P14-3-3/PELF1 complex binds PardC DNA 10 times higher than PELF1 binds to PardC DNA. Finally, our data not only represent the first report of PELF1 regulation of DNA structure formation, but also enhance our understanding of PELF1 as a DNA binding protein and PELF1/P14-3-3 interaction. |
