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首页> 外文期刊>Molecular and Cellular Biology >RIP60, a mammalian origin-binding protein, enhances DNA bending near the dihydrofolate reductase origin of replication.
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RIP60, a mammalian origin-binding protein, enhances DNA bending near the dihydrofolate reductase origin of replication.

机译：RIP60是一种哺乳动物起源结合蛋白，可增强二氢叶酸还原酶复制起点附近的DNA弯曲。

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摘要

Replication of the Chinese hamster dihydrofolate (dhfr) gene initiates near a 281-bp HaeIII fragment of stably bent DNA that binds RIP60, a 60-kDa origin-specific DNA-binding protein that has been purified from HeLa cell nuclear extract (L. Dailey, M. S. Caddle, N. Heintz, and N. H. Heintz, Mol. Cell. Biol. 10:6225-6235, 1990). Circular permutation assays showed that stable DNA bending in the dhfr origin region fragment was due to the presence of five oligo (dA)3-4 tracts, designated bend elements B1 to B5, that are spaced 10 bp apart. DNA bending directed by elements B1 to B5, as assessed by anomolous migration of DNA fragments on polyacrylamide gels, was accentuated at 4 degrees C. Bend element B5, which is in inverse orientation relative to elements B1 to B4, overlaps an ATT-rich motif that comprises the RIP60 protein-binding site. Gel mobility shift assays with circularly permuted bent DNA fragments and purified RIP60 showed that RIP60 markedly enhanced DNA bending of the dhfr origin region sequences. These results suggest that, as in many plasmids, bacteriophages, and eucaryotic viruses, mammalian DNA-binding proteins may enhance DNA bending near origins of replication during initiation of DNA synthesis.
机译：中国仓鼠二氢叶酸（dhfr）基因的复制在稳定弯曲的DNA的281 bp HaeIII片段附近开始，该片段与RIP60结合，RIP60是一种60 kDa的起源特异性DNA结合蛋白，已从HeLa细胞核提取物中纯化出来（L.Dailey ，MS Caddle，N.Heintz和NH Heintz，Mol.Cell.Biol.10：6225-6235，1990）。圆形置换分析显示dhfr起源区域片段中稳定的DNA弯曲是由于存在5个寡核苷酸（dA）3-4片段，称为弯曲元件B1至B5，其间隔为10 bp。通过DNA片段在聚丙烯酰胺凝胶上的异常迁移评估，由元素B1至B5定向的DNA弯曲在4°C时加剧。弯曲元素B5（相对于元素B1至B4呈反方向）与ATT富集重叠它包含RIP60蛋白结合位点。用圆形排列的弯曲DNA片段和纯化的RIP60进行的凝胶迁移率迁移分析表明，RIP60显着增强了dhfr起源区域序列的DNA弯曲。这些结果表明，与许多质粒，噬菌体和真核病毒一样，哺乳动物DNA结合蛋白可能会在DNA合成起始过程中增强复制起点附近的DNA弯曲。

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