TFIIF, a basal eukaryotic transcription factor, is a substrate for poly(ADP-ribosyl)ation.
TFIIF, a basal eukaryotic transcription factor, is a substrate for poly(ADP-ribosyl)ation.
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TFIIF 是一种基础真核转录因子，是聚 (ADP-核糖基) 化的底物。
DOI：
10.1042/bj3240249
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Alvarez-Gonzalez,R
中科院分区：
文献类型：
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作者：
Rawling,JM;Alvarez-Gonzalez,R
我们已经研究了一些基础的真核转录因子作为聚（ADP-核糖基）化的共价靶的敏感性。将人重组TATA结合蛋白、转录因子（TF）IIB和TFIIF（由30和74 kDa RNA聚合酶II相关蛋白RAP 30和RAP 74组成）与小牛胸腺聚（ADP-核糖）聚合酶和[32 P]NAD+在37 ℃下孵育。在十二烷基硫酸锂/PAGE和放射自显影上，观察到与RAP 30和RAP 74一致的两条放射性条带。无放射性与TATA结合蛋白或TFIIB共迁移。这种现象是依赖于切口DNA的存在，这是必不可少的聚（ADP-核糖）聚合酶活性。TFIIF的共价修饰随着孵育时间、TFIIF浓度和NAD+浓度的增加而增加。高分辨率PAGE证实，RAP 30和RAP 74相关的放射性物质是ADP-核糖聚合物。从这些观察，我们得出结论，这两个TFIIF亚基是高度特异性的共价聚（ADP-核糖基）化的基板。
We have examined the susceptibility of some of the basal eukaryotic transcription factors as covalent targets for poly(ADP-ribosyl)ation. Human recombinant TATA-binding protein, transcription factor (TF)IIB and TFIIF (made up of the 30 and 74 kDa RNA polymerase II-associated proteins RAP30 and RAP74) were incubated with calf thymus poly(ADP-ribose) polymerase and [32P]NAD+at 37 °C. On lithium dodecyl sulphate/PAGE and autoradiography, two bands of radioactivity, coincident with RAP30 and RAP74, were observed. No radioactivity co-migrated with TATA-binding protein or TFIIB. The phenomenon was dependent on the presence of nicked DNA, which is essential for poly(ADP-ribose) polymerase activity. Covalent modification of TFIIF increased with time of incubation, with increasing TFIIF concentration and with increasing NAD+concentration. High-resolution PAGE confirmed that the radioactive species associated with RAP30 and RAP74 were ADP-ribose polymers. From these observations, we conclude that both TFIIF subunits are highly specific substrates for covalent poly(ADP-ribosyl)ation.