Watching DNA Polymerase η Make a Phosphodiester Bond

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  • DNAポリメラーゼηによるリン酸ジエステル結合の形成過程の観察
  • DNA ポリメラーゼe ニ ヨル リンサン ジエステル ケツゴウ ノ ケイセイ カテイ ノ カンサツ

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Description

A number of structural and kinetic studies of DNA polymerases have proposed the catalytic mechanism of the nucleotidyl-transfer reaction. However, the actual process has never been visualized. Here we show the nucleotidyl-transfer reaction process catalyzed by human DNA polymerase η using time-resolved protein crystallography. In sequence, the nucleophile 3′-OH is deprotonated, the deoxyribose at the primer end converts from C2′-endo to C3′-endo, and the nucleophile and the α-phosphate of dATP approach each other to form the new bond. A third Mg2+ ion, which arrives with the new bond and stabilizes the intermediate state, may be an unappreciated feature of the two-metal-ion mechanism.<br>

Journal

  • Seibutsu Butsuri

    Seibutsu Butsuri 53 (5), 254-257, 2013

    The Biophysical Society of Japan General Incorporated Association

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