The association of human c-Ha-ras sequences with chromatin and nuclear proteins.
The association of human c-Ha-ras sequences with chromatin and nuclear proteins.
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人类 c-Ha-ras 序列与染色质和核蛋白的关联。
DOI：
10.1016/0006-291x(85)91668-7
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发表时间：
1985
影响因子：
3.1
通讯作者：
Smulson,M
中科院分区：
文献类型：
--
作者：
Kasid,UN;Hough,C;Thraves,P;Dritschilo,A;Smulson,M
作为了解染色质组织与癌基因表达调节之间可能关系的一步，我们研究了HeLa-S3细胞中最常激活的癌基因之一c-Ha-ras的染色质结构。这是通过分离纯化细胞核的微球菌核酸酶消化后的染色质组分（可溶性和不溶性）和探测ras序列的分布来完成的。多核小体部分通过蔗糖梯度沉降进一步解析。Southern杂交结果表明：（1）c-Ha-ras序列主要分布在裂解液中; (2)与B-珠蛋白（转录非活性）序列不同，ras-h相关染色质缺乏典型的核小体包装。此外，由于核蛋白的翻译后修饰已被认为在DNA转录和复制过程中调节核小体结构，因此还检查了在抗聚（ADP-核糖）琼脂糖上免疫分离的多核小体中的ras序列。这些数据表明，这种癌基因序列的主要类别存在于染色质中，更远离这种特定染色质修饰的位点。
As a step towards the understanding of possible relationship between chromatin organization and regulation of the oncogene expression, we have investigated the chromatin structure of one of the more frequently activated oncogenes, c-Ha-ras, in HeLa-S3 cells. This was accomplished by isolation of the chromatin fractions (soluble and insoluble) after micrococcal nuclease digestion of purified nuclei and probing for the distribution of ras sequences. The polynucleosomal fraction was further resolved by sucrose gradient sedimentation. Southern-blot hybridization of the DNA isolated from various fractions yielded following results: (1) c-Ha-ras seqences segregated predominantly in the lysate fraction. (2) Unlike theB-globin (transcriptionally inactive) sequences, ras-h associated chromatin lacked typical nucleosomal packaging. Furthermore, since post-translational modifications of nuclear proteins have been suggested to modulate the nucleosome structure during DNA transcription and replication, ras sequences, in polynucleosomes immunofractionated on anti-poly (ADP-Ribose) Sepharose were also examined. The data suggested that the major class of this oncogene sequence exists in chromatin more distal to the sites of this particular chromatin modification.