Different contributions of three zinc fingers of transcription factor Sp1 to DNA recognition: novel binding mode of N-terminal finger 1.
Nana Saegusa, Masanori Yokono, Katsuko Matsushita, Y. SUGIURA
Abstract
Nana Saegusa, Masanori Yokono, Katsuko Matsushita, Y. SUGIURA
Abstract
To clarify binding properties of the first zinc finger of Sp1, finger 1, to GC-box DNA, two-finger mutant peptides Sp1 (zf12) and Sp1 (zf23) were created and their DNA binding characteristics have been compared with those of native three-zinc finger protein Sp1. Some gel electrophoretic experiments involving DNase I footprinting and gel mobility shift assays reveal that finger 1 is not essential to sequence-specific DNA recognition, but moderately contributes to total binding affinity and overall sequence specificity by three zinc fingers of Sp1.
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To clarify binding properties of the first zinc finger of Sp1, finger 1, to GC-box DNA, two-finger mutant peptides Sp1 (zf12) and Sp1 (zf23) were created and their DNA binding characteristics have been compared with those of native three-zinc finger protein Sp1. Some gel electrophoretic experiments involving DNase I footprinting and gel mobility shift assays reveal that finger 1 is not essential to sequence-specific DNA recognition, but moderately contributes to total binding affinity and overall sequence specificity by three zinc fingers of Sp1.
Key concepts: Zinc finger, Sp1 transcription factor, DNA footprinting, Zinc finger nuclease, RING finger domain, DNA-binding protein, DNA, Transcription factor