DNA結合蛋白
DNA結合蛋白(DNA-binding protein)是指能透過DNA結合結構域(DBD)與單股或雙股DNA結合的一類蛋白質[1][2][3]。一般來說,能與特異DNA序列結合的DNA結合蛋白主要經由與B-DNA(生物體內DNA一般都是B-DNA形式)的大溝結合來識別DNA序列。因為B-DNA的大溝區域能容許更多蛋白質上的官能基靠近並與之進行相互作用[4][5]。
蛋白質與DNA的相互作用
編輯蛋白質與DNA的結合受到pH值、溫度、離子強度、電場,以及大分子擁擠等多種條件的影響,具體分為特異性與非特異性兩種。非特異性的結合指蛋白質與DNA的結合不依賴特測序列,而特異性的結合則只會發生在特定的DNA序列上[6][7]。在Cas9/CRSIPER系統出現前作為主流基因編輯工具的鋅指蛋白以及TALENs都是依賴特異性的DNA蛋白間結合工作的[8]。
檢測方法
編輯較早期的檢測蛋白與DNA間相互作用的方法[9]包括凝膠遷移實驗(EMSA)[10][11]、DNA-蛋白質相互作用ELISA實驗(DPI-ELISA)[12]、DNA酶足跡法(DNase footprinting assay) [13]等等。而染色質免疫沉澱法(ChIP)則是目前較為通行的檢測體內DNA與蛋白質相互作用的方法[14]。
非特異性DNA結合蛋白
編輯最常見的一種非特異性DNA結合蛋白是真核生物染色質中與DNA結合的組蛋白。組蛋白主要是通過所帶的正電荷與帶負電的DNA結合,所以組蛋白與DNA之間的結合不依賴DNA上的特測序列。這也符合組蛋白的生物學功能,因為組蛋白需要與各種不同的DNA片段結合形成染色質的基本組成單位核小體[15][16]。另外,真核生物染色質上能夠使DNA鏈發生彎曲的高遷移率族(HMG)蛋白與DNA之間的結合也是非特異性的[17][18][19]。
特異性DNA結合蛋白
編輯很多特異性DNA結合蛋白屬於轉錄因子。轉錄因子依賴其特有的DNA結合結構域與DNA上的特測序列結合。一般來說,轉錄因子在與DNA結合後,會招募酶對DNA或組蛋白進行修飾,進而調控基因的表達水平[20][21]。
參見
編輯參考文獻
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