ISOLATION AND CHARACTERIZATION OF HUMAN CDNA CLONES ENCODING A HIGH MOBILITY GROUP BOX PROTEIN THAT RECOGNIZES STRUCTURAL DISTORTIONS TO DNA CAUSED BY BINDING OF THE ANTICANCER AGENT CISPLATIN
ISOLATION AND CHARACTERIZATION OF HUMAN CDNA CLONES ENCODING A HIGH MOBILITY GROUP BOX PROTEIN THAT RECOGNIZES STRUCTURAL DISTORTIONS TO DNA CAUSED BY BINDING OF THE ANTICANCER AGENT CISPLATIN
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DOI：
10.1073/pnas.89.6.2307
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发表时间：
1992-03-15
影响因子：
11.1
通讯作者：
LIPPARD, SJ
中科院分区：
文献类型：
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作者：
BRUHN, SL;PIL, PM;LIPPARD, SJ
编码结构特异性识别蛋白 SSRP1 的人类 cDNA 克隆已被分离和表征，该蛋白特异性结合顺铂修饰的 DNA。 SSRP1 基因定位于人类染色体 11q12。通过使用之前描述的部分长度 cDNA 获得的 cDNA 克隆预测出一个 81 kDa 的蛋白质，其中包含几个高电荷结构域和一段 75 个氨基酸，与高迁移率基团 (HMG) 蛋白质 HMG1 的一部分具有 47% 的一致性。该 HMG 盒很可能构成顺铂修饰 DNA 的结构识别元件，可能的识别基序是局部双链体解旋并向主沟弯曲，该主沟在链内 cis-{Pt(NH3)2}2+ d(GpG) 和 d(ApG) 交联形成时发生。尽管 HMG-box 蛋白质家族成员的 DNA 识别特性已根据其序列特异性进行了表征，但目前的工作表明，具有该结构域的蛋白质也可以识别特定的 DNA 结构。这里描述的 Pt-DNA SSRP 是最近鉴定的小鼠蛋白的人类同源物，该蛋白结合重组信号序列 [Shirakata, M.、Huppi, K.、Usuda, S.、Okazaki, K.、Yoshida, K. & Sakano, H. (1991) Mol.细胞。生物。 11, 4528-4536]。这些序列被假定形成茎环结构，进一步暗示 DNA 中的局部弯曲和解旋可以作为 HMG-box 蛋白的识别目标。多种组织和顺铂耐药细胞系中的表达分析以及顺铂无法在 HeLa 细胞中诱导该信息，反对 SSRP1 mRNA 水平与细胞对药物反应之间的直接联系。
Human cDNA clones encoding a structure-specific recognition protein, SSRP1, that binds specifically to DNA modified with cisplatin have been isolated and characterized. The SSRP1 gene maps to human chromosome 11q12. The cDNA clones, obtained by using partial-length cDNAs described previously, predict an 81-kDa protein containing several highly charged domains and a stretch of 75 amino acids 47% identical to a portion of the high mobility group (HMG) protein HMG1. This HMG box most likely constitutes the structure recognition element for cisplatin-modified DNA, with the probable recognition motif being the local duplex unwinding and bending toward the major groove that occurs upon formation of intra-strand cis-{Pt(NH3)2}2+ d(GpG) and d(ApG) cross-links. Although the DNA recognition properties of members of the HMG-box family of proteins have been characterized with respect to their sequence specificity, the present work demonstrates that proteins with this domain can recognize particular DNA structures as well. The Pt-DNA SSRP described here is the human homolog of a recently identified mouse protein that binds to recombination signal sequences [Shirakata, M., Huppi, K., Usuda, S., Okazaki, K., Yoshida, K. & Sakano, H. (1991) Mol. Cell. Biol. 11, 4528-4536]. These sequences have been postulated to form stem-loop structures, further implicating local bends and unwinding in DNA as a recognition target for HMG-box proteins. Expression analysis in a variety of tissues and cisplatin-resistant cell lines and the inability of cisplatin to induce the message in HeLa cells argue against a direct link between SSRP1 mRNA levels and the response of cells to the drug.