Uchida K, Hanai S, Ishikawa K, Ozawa Y, Uchida M, Sugimura T, Miwa M
Department of Biochemistry, University of Tsukuba, Ibaraki, Japan.
Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3481-5. doi: 10.1073/pnas.90.8.3481.
We have isolated cDNA clones for a Drosophila poly(ADP-ribose) polymerase (PARP; EC 2.4.2.30) by screening a lambda gt11 cDNA library with a Drosophila partial cDNA fragment. The Drosophila PARP probe was obtained by the polymerase chain reaction with heterologous primers deduced from conserved amino acids in the mammalian, chicken, amphibian, and fish sequences. The Drosophila PARP mRNA is 3.2 kb in length and is expressed in the early stages of development. The PARP protein of 994 amino acids contains two zinc-finger motifs and an NAD-binding motif, which are conserved among different species. Interestingly, the heptad leucine repeat in an alpha-helix was found in Drosophila PARP. Alignments of the auto-modification domains of various species showed the repeated hydrophobic amino acids on the same face of the helix that make the coiled-coil configuration in the mammalian and chicken sequences. The presence of a leucine-zipper motif in the auto-modification domain suggests that this motif might be responsible for protein-protein interaction between PARP and physiological acceptors. PARP may have novel functions, possibly involving its homo- and/or heterodimerization with other nuclear leucine-zipper proteins and its regulation by ADP-ribosylation.
我们通过用果蝇部分cDNA片段筛选λgt11 cDNA文库,分离出了果蝇多聚(ADP - 核糖)聚合酶(PARP;EC 2.4.2.30)的cDNA克隆。果蝇PARP探针是通过聚合酶链反应获得的,所用异源引物是根据哺乳动物、鸡、两栖动物和鱼类序列中保守氨基酸推导出来的。果蝇PARP mRNA长度为3.2 kb,在发育早期表达。994个氨基酸的PARP蛋白包含两个锌指基序和一个NAD结合基序,这些在不同物种中是保守的。有趣的是,在果蝇PARP中发现了α - 螺旋中的七肽亮氨酸重复序列。各种物种自身修饰结构域的比对显示,螺旋同一面上存在重复的疏水氨基酸,这在哺乳动物和鸡的序列中形成了卷曲螺旋结构。自身修饰结构域中亮氨酸拉链基序的存在表明,该基序可能负责PARP与生理受体之间的蛋白质 - 蛋白质相互作用。PARP可能具有新功能,可能涉及其与其他核亮氨酸拉链蛋白的同型和/或异型二聚化以及其通过ADP - 核糖基化的调节。