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首页> 外文期刊>Molecular and Cellular Biology >RNA polymerase III transcription from the human U6 and adenovirus type 2 VAI promoters has different requirements for human BRF, a subunit of human TFIIIB.
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RNA polymerase III transcription from the human U6 and adenovirus type 2 VAI promoters has different requirements for human BRF, a subunit of human TFIIIB.

机译：从人U6和2型腺病毒VAI启动子转录RNA聚合酶III对人BRF（人TFIIIB的一个亚基）有不同的要求。

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Mammalian TFIIIB can be separated into two fractions required for transcription of the adenovirus type 2 VAI gene, which have been designated 0.38M-TFIIIB and 0.48M-TFIIIB. While 0.48M-TFIIIB has not been characterized, 0.38M-TFIIIB corresponds to a TBP-containing complex. We describe here the purification of this complex, which consists of TBP and a closely associated polypeptide of 88 kDa, and the isolation of a cDNA corresponding to the 88-kDa polypeptide. The predicted protein sequence reveals that the 88-kDa polypeptide corresponds to a human homolog of the Saccharomyces cerevisiae BRF protein, a subunit of yeast TFIIIB. Human BRF (hBRF) probably corresponds to TFIIIB90, a protein previously cloned by Wang and Roeder (Proc. Natl. Acad. Sci. USA 92:7026-7030, 1995), although its predicted amino acid sequence differs from that reported for TFIIIB90 over a stretch of 67 amino acids as a result of frameshifts. Immunodepletion of more than 90 to 95% of the hBRF present in a transcription extract severely debilitates transcription from the tRNA-type VAI promoter but does not affect transcription from the TATA box-containing human U6 promoter, suggesting that the 0.38M-TFIIIB complex, and perhaps hBRF as well, is not required for U6 transcription.
机译：哺乳动物TFIIIB可分为2型腺病毒VAI基因转录所需的两个部分，分别称为0.38M-TFIIIB和0.48M-TFIIIB。虽然没有表征0.48M-TFIIIB，但是0.38M-TFIIIB对应于含有TBP的复合物。我们在此描述了该复合物的纯化，该复合物由TBP和88 kDa的紧密相关多肽组成，以及与88 kDa多肽相对应的cDNA的分离。预测的蛋白质序列揭示了88-kDa多肽对应于酿酒酵母BRF蛋白（酵母TFIIIB的一个亚基）的人类同源物。人BRF（hBRF）可能对应于TFIIIB90，这是先前由Wang和Roeder（Proc。Natl。Acad。Sci。USA 92：7026-7030，1995）克隆的蛋白质，尽管其预测的氨基酸序列与TFIIIB90报道的氨基酸序列不同由于移码而延伸的67个氨基酸。转录提取物中存在超过90％至95％的hBRF的免疫耗竭会严重破坏tRNA型VAI启动子的转录，但不会影响来自包含TATA盒的人U6启动子的转录，这表明0.38M-TFIIIB复合物U6转录可能并不需要hBRF。

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