Kochiwa Hiromi, Itaya Mitsuhiro, Tomita Masaru, Kanai Akio
Institute for Advanced Biosciences, Keio University, Tsuruoka, Japan.
FEBS J. 2006 Jan;273(2):420-9. doi: 10.1111/j.1742-4658.2005.05082.x.
Ribonuclease H1 (RNase H1) is a widespread enzyme found in a range of organisms from viruses to humans. It is capable of degrading the RNA moiety of DNA-RNA hybrids and requires a bivalent ion for activity. In contrast with most eukaryotes, which have one gene encoding RNase H1, the activity of which depends on Mg(2+) ions, Caenorhabditis elegans has four RNase H1-related genes, and one of them has an isoform produced by alternative splicing. However, little is known about the enzymatic features of the proteins encoded by these genes. To determine the differences between these enzymes, we compared the expression patterns of each RNase H1-related gene throughout the development of the nematode and the RNase H activities of their recombinant proteins. We found gene-specific expression patterns and different enzymatic features. In particular, besides the enzyme that displays the highest activity in the presence of Mg(2+) ions, C. elegans has another enzyme that shows preference for Mn(2+) ion as a cofactor. We characterized this Mn(2+)-dependent RNase H1 for the first time in eukaryotes. These results suggest that there are at least two types of RNase H1 in C. elegans depending on the developmental stage of the organism.
核糖核酸酶H1(RNase H1)是一种广泛存在于从病毒到人类等一系列生物体中的酶。它能够降解DNA-RNA杂交体中的RNA部分,并且其活性需要二价离子。与大多数真核生物不同,大多数真核生物有一个编码RNase H1的基因,其活性依赖于Mg(2+)离子,秀丽隐杆线虫有四个与RNase H1相关的基因,其中一个有通过可变剪接产生的同种型。然而,对于这些基因所编码蛋白质的酶学特性知之甚少。为了确定这些酶之间的差异,我们比较了每个与RNase H1相关的基因在秀丽隐杆线虫整个发育过程中的表达模式及其重组蛋白的RNase H活性。我们发现了基因特异性的表达模式和不同的酶学特性。特别是,除了在Mg(2+)离子存在下表现出最高活性的酶之外,秀丽隐杆线虫还有另一种酶,它表现出对Mn(2+)离子作为辅因子的偏好。我们首次在真核生物中对这种依赖Mn(2+)的RNase H1进行了表征。这些结果表明,根据生物体的发育阶段,秀丽隐杆线虫中至少存在两种类型的RNase H1。