Luque A E, Benedetto J P, Castroviejo M
REGER, EP 630, IBGC-CNRS, Bordeaux, France.
Plant Mol Biol. 1998 Nov 1;38(4):647-54. doi: 10.1023/a:1006052213299.
We have previously described a low-molecular-weight DNA polymerase (52 kDa) from wheat embryo: DNA polymerase CI (pol CI). This enzyme shares some biochemical properties with animal DNA polymerase beta (pol beta). In this report, we analyse pol CI in wheat embryo germination. Immunodetection and measurement of the enzyme activity show that wheat pol CI remains at a constant level during germination, whereas dramatic changes of the replicative DNA polymerase A and B activities were previously reported. We observe that the level of pol CI in physiological conditions (embryo germination and dividing cell culture) is in agreement with a pol beta-type DNA polymerase. By microsequencing of the electroblotted 52 kDa polypeptide, we determined the sequence of a dodecapeptide from the N-terminal region. A comparative analysis of the N-terminal pol CI peptide with some mammalian pol beta sequences shows a clear homology with helix 1 of the N-terminal ssDNA domain (residues 15 to 26) of the rat pol beta. Thus, the helical structure of this region should be conserved in the wheat peptide. This represents the first evidence of a partial primary structure of a beta-type DNA polymerase in plants.
我们之前曾描述过从小麦胚中分离出的一种低分子量DNA聚合酶(52 kDa):DNA聚合酶CI(pol CI)。这种酶与动物DNA聚合酶β(pol β)具有一些生化特性。在本报告中,我们分析了小麦胚萌发过程中的pol CI。酶活性的免疫检测和测量结果表明,在萌发过程中小麦pol CI的水平保持恒定,而之前报道过复制性DNA聚合酶A和B的活性有显著变化。我们观察到,在生理条件下(胚萌发和分裂细胞培养)pol CI的水平与pol β型DNA聚合酶一致。通过对电印迹的52 kDa多肽进行微量测序,我们确定了来自N端区域的一个十二肽的序列。对N端pol CI肽与一些哺乳动物pol β序列的比较分析表明,它与大鼠pol β的N端单链DNA结构域(第15至26位残基)的螺旋1有明显的同源性。因此,该区域的螺旋结构在小麦肽中应该是保守的。这是植物中β型DNA聚合酶部分一级结构的首个证据。