Heller-Uszynska Katarzyna, Schnippenkoetter Wendelin, Kilian Andrzej
CAMBIA: Center for the Application of Molecular Biology to International Agriculture, GPO Box 3200, Canberra, ACT 2601, Australia.
Plant J. 2002 Jul;31(1):75-86. doi: 10.1046/j.1365-313x.2001.01337.x.
Plant chromosomes terminate in telomeres as in other eukaryotes. Telomeres are vital to genome stability and their malfunctioning is lethal. One of the core components of the telomere complex is telomerase. The enzyme activity depends on RNA (TER) and reverse transcriptase (TERT) subunits. We describe here the isolation, sequencing and characterization of the telomerase reverse transcriptase catalytic subunit from the monocot plant Oryza sativa L. (OsTERT). A single copy of this gene is present in the rice genome. The protein predicted from the OsTERT sequence has all the signature motifs of the TERT family members. Our data indicate that rice telomerase activity is developmentally regulated and is high in in vitro tissue and cell culture. However, steady-state transcript levels of the TERT gene do not seem to correlate with enzyme activity. Northern and RT-PCR analyses of the OsTERT gene transcript profile show multiple differentially spliced transcripts in both telomerase-positive and telomerase-negative tissues. Based on quantitative analysis of these transcripts, we speculate that the overall balance between the quantities of particular alternatively spliced transcripts may determine whether the TERT protein(s) is active or not. The diversity of splicing variants detected suggests that, as recently discovered for mammalian TERT proteins, rice TERT protein variants may perform functions other than telomere maintenance.
与其他真核生物一样,植物染色体的末端是端粒。端粒对于基因组稳定性至关重要,其功能异常是致命的。端粒复合体的核心成分之一是端粒酶。该酶的活性取决于RNA(TER)和逆转录酶(TERT)亚基。我们在此描述了从单子叶植物水稻(Oryza sativa L.)中分离、测序和鉴定端粒酶逆转录酶催化亚基(OsTERT)的过程。水稻基因组中存在该基因的单拷贝。从OsTERT序列预测的蛋白质具有TERT家族成员的所有特征基序。我们的数据表明,水稻端粒酶活性受到发育调控,在体外组织和细胞培养中活性较高。然而,TERT基因的稳态转录水平似乎与酶活性无关。对OsTERT基因转录谱的Northern和RT-PCR分析表明,在端粒酶阳性和端粒酶阴性组织中均存在多个差异剪接的转录本。基于对这些转录本的定量分析,我们推测特定可变剪接转录本数量之间的总体平衡可能决定TERT蛋白是否具有活性。检测到的剪接变体的多样性表明,正如最近在哺乳动物TERT蛋白中发现的那样,水稻TERT蛋白变体可能具有除维持端粒之外的其他功能。