Enhanced processivity of nuclear matrix bound DNA polymerase alpha from regenerating rat liver.
Enhanced processivity of nuclear matrix bound DNA polymerase alpha from regenerating rat liver.
复制标题
再生大鼠肝脏中核基质结合 DNA 聚合酶 α 的持续合成能力增强。
DOI：
10.1021/bi00392a020
复制
发表时间：
1987
期刊：
影响因子：
2.9
通讯作者：
Berezney,R
中科院分区：
文献类型：
--
作者：
Tubo,RA;Martelli,AM;Berezney,R
Ross A. Tubo，** Alberto M. Martelli，and罗纳德Berezney* Department of Biological Sciences，州立大学of纽约布法罗，布法罗，纽约14260 1987年4月10日接收摘要：通过测定再生大鼠肝细胞核基质结合复制组装体的持续合成能力，（聚合酶与DNA模板的一次生产性结合事件后添加的核苷酸的平均数）的核基质结合的DNA聚合酶a与作为模板引物的聚（dT）* 寡聚（A）10的比较。基质结合的聚合酶的平均持续合成能力（28.4个核苷酸）是几倍高于散装核DNA聚合酶的活性提取核基质制备过程中（8.9个核苷酸）。ATP在1 mM显着增强的活性和持续合成能力的基质结合的聚合酶，但不是相应的盐溶性酶。大部分ATP依赖性活性和持续合成能力增强由100 μ M ATP完成，并且包括范围高达完整模板长度（1000-1200个核苷酸）的产物。净ATP刺激的聚合酶活性的平均持续合成能力超过80个核苷酸，几乎所有的DNA产物> 50个核苷酸。通过超声处理释放核基质结合的DNA聚合酶a导致ATP刺激活性的丧失和相应的持续合成能力降低到与盐溶性聚合酶（6.8个核苷酸）相似的水平。所有核苷二磷酸和三磷酸与ATP一样有效。通过不可水解的ATP类似物腺苷5/-O-（3-硫代三磷酸）、5 '-腺苷酰亚胺二磷酸和腺苷-O-（1-硫代三磷酸）对活性和持续合成能力的刺激进一步表明，ATP的水解不是发生增强所必需的。核苷一磷酸、ATP类似物P1，P4-双（5/-腺苷）四磷酸和5 '-腺苷酰亚甲基二磷酸、ADP不可水解类似物腺苷5'-O-（2-硫代二磷酸）和焦磷酸无法增强活性或持续合成能力，表明了核苷酸激活的一定程度的特异性。
Ross A. Tubo,** Alberto M. Martelli, and Ronald Berezney* Department of Biological Sciences, State University of New York at Buffalo, Buffalo, New York 14260 Received April 10, 1987 abstract: Translocation of DNA duringin vitroDNA synthesis on nuclear matrix bound replicational assemblies from regeneratingrat liver was determined by measuring the processivity (average number of nucleotides added following one productive binding event of the polymerase to the DNA template) of nuclear matrix bound DNA polymerase a with poly (dT)* oligo (A) 10 as template primer. The matrix-bound polymerase had an average processivity (28.4 nucleotides) that was severalfold higher than the bulk nuclear DNA polymerase a activity extracted during nuclear matrix preparation (8.9 nucleotides). ATP at 1 mM markedly enhanced the activity and processivity of the matrix-bound polymerase but not the corresponding salt-soluble enzyme. The majority of the ATP-dependent activity and processivity enhancement was completed by 100 juM ATP and included products ranging up to full template length (1000-1200 nucleotides). Average processivity of the net ATP-stimulated polymerase activity exceeded 80 nucleotides with virtually all the DNA products> 50 nucleotides. Release of nuclear matrix bound DNA polymerase a by sonication resulted in a loss of ATP stimulation of activity and a corresponding decrease in processivity to a level similar to that of the salt-solublepolymerase (6.8 nucleotides). All nucleoside di-and triphosphates were as effective as ATP. Stimulation of both activity and processivity by the nonhydrolyzable ATP analogues adenosine 5/-0-(3-thiotriphosphate), 5'-adenylyl imidodiphosphate, and adenosine^-(^-(l-thiotriphosphate) further suggested that the hydrolysis of ATP is not requiredfor enhancement to occur. A degree of specificity of nucleotide activation was indicated by the inability of nucleoside monophosphates, the ATP analogues P1, P4-bis (5/-adenosyl) tetraphosphate and 5'-adenylyl methylenediphosphate, the ADP-nonhydrolyzable analogue adenosine 5'-0-(2-thiodiphosphate), and pyrophosphate to enhance either activity or processivity.