Kumar A, Wilson S H
Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Biochemistry. 1990 Dec 4;29(48):10717-22. doi: 10.1021/bi00500a001.
A1 is a major core protein of the mammalian hnRNP complex, and as a purified protein of approximately 34 kDa, A1 is a strong single-stranded nucleic acid binding protein. Several lines of evidence suggest that the protein is organized in discrete domains consisting of an N-terminal segment of approximately 22 kDa and a C-terminal segment of approximately 12 kDa. Each of these domains as a purified fragment is capable of binding to both ssDNA and RNA. We report here that A1 and its C-terminal domain fragment are capable of potent strand-annealing activity for base-pair complementary single-stranded polynucleotides of both RNA and DNA. This effect is not stimulated by ATP. Compared with A1 and the C-terminal fragment, the N-terminal domain fragment has negligible annealing activity. These results indicate that A1 has biochemical activity consistent with a strand-annealing role in relevant reactions, such as pre-mRNA splicing.
A1是哺乳动物hnRNP复合物的一种主要核心蛋白,作为一种约34 kDa的纯化蛋白,A1是一种强大的单链核酸结合蛋白。多项证据表明,该蛋白由离散结构域组成,包括一个约22 kDa的N端片段和一个约12 kDa的C端片段。这些结构域中的每一个作为纯化片段都能够与单链DNA和RNA结合。我们在此报告,A1及其C端结构域片段对RNA和DNA的碱基对互补单链多核苷酸具有强大的链退火活性。这种效应不受ATP的刺激。与A1和C端片段相比,N端结构域片段的退火活性可忽略不计。这些结果表明,A1具有与相关反应(如前体mRNA剪接)中的链退火作用一致的生化活性。