Biswas S B, Biswas E E
Division of Endocrinology, University of Maryland School of Medicine, Baltimore 21201.
Mol Cell Biol. 1990 Feb;10(2):810-5. doi: 10.1128/mcb.10.2.810-815.1990.
We have analyzed various autonomously replicating sequences (ARSs) in yeast nuclear extract with ARS-specific synthetic oligonucleotides. The EI oligonucleotide sequence, which is derived from HMRE-ARS, and the F1 oligonucleotide sequence, which is derived from telomeric ARS120, appeared to bind to the same cellular factor with high specificity. In addition, each of these oligonucleotides was a competitive inhibitor of the binding of the other. Binding of the ARS binding factor (ABF) to either of these oligonucleotides was inhibited strongly by plasmids containing ARS1 and telomeric TF1-ARS. DNase I footprinting analyses with yeast nuclear extract showed that EI and F1 oligonucleotides eliminated protection of the binding site of ARS binding factor I (ABFI) in domain B of ARS1. Sequence analyses of various telomeric (ARS120 and TF1-ARS) and nontelomeric ARSs (ARS1 and HMRE-ARS) showed the presence of consensus ABFI binding sites in the protein binding domains of all of these ARSs. Consequently, the ABFI and ABFI-like factors bind to these domain B-like sequences in a wide spectrum of ARSs, both telomeric and nontelomeric.
我们用ARS特异性合成寡核苷酸分析了酵母核提取物中的各种自主复制序列(ARSs)。源自HMRE - ARS的EI寡核苷酸序列和源自端粒ARS120的F1寡核苷酸序列,似乎以高特异性结合同一细胞因子。此外,这些寡核苷酸中的每一个都是另一个结合的竞争性抑制剂。含有ARS1和端粒TF1 - ARS的质粒强烈抑制ARS结合因子(ABF)与这些寡核苷酸中任何一个的结合。用酵母核提取物进行的DNase I足迹分析表明,EI和F1寡核苷酸消除了ARS1结构域B中ARS结合因子I(ABFI)结合位点的保护。对各种端粒(ARS120和TF1 - ARS)和非端粒ARSs(ARS1和HMRE - ARS)的序列分析表明,在所有这些ARSs的蛋白质结合结构域中存在共有ABFI结合位点。因此,ABFI和ABFI样因子在广泛的ARSs(包括端粒和非端粒)中与这些结构域B样序列结合。