Activation of p53 Facilitates the Target Search in DNA by Enhancing the Target Recognition Probability.
Tumor suppressor p53 binds to the target in a genome and regulates the expression of downstream genes. p53 searches for the target by combining three-dimensional diffusion and one-dimensional sliding along the DNA. To examine the regulation mechanism of the target binding, we constructed the pseudo-wild type (pseudo-WT), activated (S392E), and inactive (R248Q) mutants of p53 and observed their target binding in long DNA using single-molecule fluorescence imaging. The pseudo-WT sliding along the DNA showed many pass events over the target and possessed target recognition probability (TRP) of 7±2%. The TRP increased to 18±2% for the activated mutant but decreased to 0% for the inactive mutant. Furthermore, the fraction of the target binding by the one-dimensional sliding among the total binding events increased from 63±9% for the pseudo-WT to 87±2% for the activated mutant. Control of TRP upon activation, as demonstrated here for p53, might be a general activation mechanism of transcription factors.
Institute for Multidisciplinary Research for Advanced Materials, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai 980-8577, Japan; Department of Chemistry, Graduate School of Science, Tohoku University, Sendai 980-8578, Japan.
Yuji
Y
Itoh
Institute for Multidisciplinary Research for Advanced Materials, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai 980-8577, Japan; Department of Chemistry, Graduate School of Science, Tohoku University, Sendai 980-8578, Japan.
Agato
A
Murata
Institute for Multidisciplinary Research for Advanced Materials, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai 980-8577, Japan.
Seiji
S
Sakamoto
Department of Microbial Resources, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan.
Kei
K
Nanatani
Institute for Multidisciplinary Research for Advanced Materials, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai 980-8577, Japan.
Takehiko
T
Wada
Institute for Multidisciplinary Research for Advanced Materials, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai 980-8577, Japan; Department of Chemistry, Graduate School of Science, Tohoku University, Sendai 980-8578, Japan. Electronic address: st@tagen.tohoku.ac.jp.
Satoshi
S
Takahashi
Institute for Multidisciplinary Research for Advanced Materials, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai 980-8577, Japan; Department of Chemistry, Graduate School of Science, Tohoku University, Sendai 980-8578, Japan. Electronic address: kamagata@tagen.tohoku.ac.jp.
Kiyoto
K
Kamagata
D001665:Binding Sites / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D011336:Probability; D011485:Protein Binding / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53; D004247:DNA
mutation; p53; single molecule; target binding; transcription factor