Török T, Gorjánácz M, Bryant P J, Kiss I
Institute of Genetics, Biological Research Center of the Hungarian Academy of Sciences, Szeged H-6701, Hungary.
Nucleic Acids Res. 2000 Sep 15;28(18):3551-7. doi: 10.1093/nar/28.18.3551.
The proliferation disrupter (prod) gene of Drosophila melanogaster encodes a novel protein associated with centromeric chromosomal regions that is required for chromatin condensation and cell viability. We have examined the binding of the Prod protein to DNA in vitro. Co-immunoprecipitation experiments demonstrate that Prod is a DNA-binding protein that specifically recognizes the 10 bp AGAATAACAT satellite repeat of D.melanogaster. Footprinting experiments show that the protein interacts with a 5-8 bp target sequence in each 10 bp repeat and suggest that it can mediate condensation of this satellite into a superhelix. Gel retardation experiments indicate that Prod does not have a well defined DNA-binding domain and it binds the satellite in a co-operative manner, probably forming Prod multimers. Since Prod localizes to both heterochromatin and euchromatin in vivo, we discuss the possibility that the ability of pre-existing euchromatic proteins to bind DNA in a co-operative manner, might be a prerequisite of satellite compaction and satellite amplification, thereby providing a basic factor in heterochromatin evolution.
黑腹果蝇的增殖干扰基因(prod)编码一种与着丝粒染色体区域相关的新型蛋白质,该蛋白质是染色质浓缩和细胞活力所必需的。我们已经在体外检测了Prod蛋白与DNA的结合。免疫共沉淀实验表明,Prod是一种DNA结合蛋白,它能特异性识别黑腹果蝇的10bp AGAATAACAT卫星重复序列。足迹实验表明,该蛋白与每10bp重复序列中的一个5-8bp靶序列相互作用,并表明它可以介导这种卫星序列浓缩成超螺旋。凝胶阻滞实验表明,Prod没有明确界定的DNA结合结构域,它以协同方式结合卫星序列,可能形成Prod多聚体。由于Prod在体内定位于异染色质和常染色质,我们讨论了预先存在的常染色质蛋白以协同方式结合DNA的能力可能是卫星序列压缩和卫星序列扩增的先决条件,从而为异染色质进化提供一个基本因素的可能性。