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PCR Cloning and Subcloning> | ... ref. 23. A complementary DNA encoding E2F8 DBD, containing N-terminal thioredoxin and a 6 × His-tag, optimized for expression in E. coli was purchased from GenScript and subcloned into the pETG-20A vector. The construct ... | Get A Quote |
The mammalian cell cycle is controlled by the E2F family of transcription factors. Typical E2Fs bind to DNA as heterodimers with the related dimerization partner (DP) proteins, whereas the atypical E2Fs, E2F7 and E2F8 contain two DNA-binding domains (DBDs) and act as repressors. To understand the mechanism of repression, we have resolved the structure of E2F8 in complex with DNA at atomic resolution. We find that the first and second DBDs of E2F8 resemble the DBDs of typical E2F and DP proteins, respectively. Using molecular dynamics simulations, biochemical affinity measurements and chromatin immunoprecipitation, we further show that both atypical and typical E2Fs bind to similar DNA sequences in vitro and in ... More