Chemistry Department, Lomonosov Moscow State University.
Acta Naturae. 2012 Jan;4(1):70-3.
Telomerase is a ribonucleoprotein, the main function of which is to synthesize telomeres, i.e. repetitive sequences which are localized at the ends of eukaryotic chromosomes. Telomerase maintains the stability of the genome in eukaryotic cells by replicating chromosomal ends. The structural and functional investigation of the telomerase complex is significantly restricted due to difficulties connected with the isolation of its main catalytic subunit in recombinant form. Herein, we describe a method developed for the isolation of the recombinant telomerase reverse transcriptase from thermotolerant yeastHansenula polymorpha. A functional test performed for the isolated protein and the RNA/DNA duplex, simulating the interaction of telomerase RNA and telomere, reveals that the isolated catalytic subunit of telomerase possesses limited reverse transcriptase activity.
端粒酶是一种核糖核蛋白,其主要功能是合成端粒,即定位于真核染色体末端的重复序列。端粒酶通过复制染色体末端来维持真核细胞基因组的稳定性。由于难以以重组形式分离其主要催化亚基,因此对端粒酶复合物的结构和功能研究受到了很大限制。在此,我们描述了一种从耐热酵母海栖热袍菌中分离重组端粒酶逆转录酶的方法。对分离的蛋白质和 RNA/DNA 双链体(模拟端粒酶 RNA 与端粒的相互作用)进行的功能测试表明,分离的端粒酶催化亚基具有有限的逆转录酶活性。