Purification and properties of the Rous sarcoma virus internal enhancer binding factor.

L Karnitz; D Poon; P A Weil; R Chalkley

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Purification and properties of the Rous sarcoma virus internal enhancer binding factor.

机译：劳斯肉瘤病毒内部增强子结合因子的纯化和性质。

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The internal enhancer binding factor (IBF) that specifically binds sequences within the gag gene internal enhancer of Rous sarcoma virus Schmidt-Ruppin A was purified to near homogeneity from BHK cells. The polypeptides that constituted IBF DNA-binding activity were identified by sodium dodecyl sulfate-polyacrylamide gel analysis. As isolated from BHK cells, IBF consisted of two different but related polypeptides. One (IBF alpha) had a molecular weight of 40,000; the other (IBF beta) had a molecular weight of 20,000 and appeared to be a proteolytic product of IBF alpha. The site within the gag gene to which IBF bounds in vitro (internal enhancer site 2; nucleotides 856 to 878 of the Rous sarcoma virus genome) were demonstrated to function as a cis-acting transcriptional stimulatory element both in vivo and in vitro. By using HeLa cell nuclear transcription extracts, purified IBF was found to function as a trans-acting transcription factor that stimulated transcription in vitro. Purified IBF was also demonstrated to be very similar to EBP20 (K. Carlberg, T. A. Ryden, and K. Beemon, J. Virol. 62:1617-1624, 1988), and it may well belong to the same family of DNA-binding proteins.

机译：从BHK细胞中纯化特异性结合Rous肉瘤病毒Schmidt-Ruppin A的gag基因内部增强子内序列的内部增强子结合因子（IBF）。通过十二烷基硫酸钠-聚丙烯酰胺凝胶分析鉴定了构成IBF DNA结合活性的多肽。从BHK细胞中分离出来的IBF由两种不同但相关的多肽组成。一种（IBFα）的分子量为40,000。另一个（IBF beta）的分子量为20,000，似乎是IBFα的蛋白水解产物。已证明gaf基因中与IBF体外结合的位点（内部增强子位点2;劳斯肉瘤病毒基因组的核苷酸856至878）在体内和体外均起顺式作用的转录刺激元件的作用。通过使用HeLa细胞核转录提取物，发现纯化的IBF可以充当反式转录因子，在体外刺激转录。还证明了纯化的IBF与EBP20非常相似（K.Carlberg，TA Ryden，和K.Beemon，J.Virol.62：1617-1624，1988），并且它很可能属于相同的DNA结合家族。蛋白质。

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《Molecular and Cellular Biology》 |1989年第5期|共11页
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L Karnitz; D Poon; P A Weil; R Chalkley;
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6. Purification and properties of the Rous sarcoma virus internal enhancer binding factor. [O] . L Karnitz, D Poon, P A Weil, 1989

机译：劳斯肉瘤病毒内部增强子结合因子的纯化和性质。
7. Purification and properties of the Rous sarcoma virus internal enhancer binding factor. [O] . Karnitz, L, Poon, D, Weil, P A, 1989

机译：劳斯肉瘤病毒内部增强子结合因子的纯化和性质。

Purification and properties of the Rous sarcoma virus internal enhancer binding factor.

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