Bautista-España Dolores, Anastacio-Marcelino Estela, Horta-Valerdi Guillermo, Celestino-Montes Antonio, Kojic Milorad, Negrete-Abascal Erasmo, Reyes-Cervantes Hortensia, Vázquez-Cruz Candelario, Guzmán Plinio, Sánchez-Alonso Patricia
Centro de Investigaciones en Ciencias Microbiológicas, Instituto de Ciencias, Benemérita Universidad Autónoma de Puebla, Puebla, Puebla, Mexico.
Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, Serbia.
PLoS One. 2014 Oct 9;9(10):e109981. doi: 10.1371/journal.pone.0109981. eCollection 2014.
In this study, we investigated the reverse transcriptase subunit of telomerase in the dimorphic fungus Ustilago maydis. This protein (Trt1) contains 1371 amino acids and all of the characteristic TERT motifs. Mutants created by disrupting trt1 had senescent traits, such as delayed growth, low replicative potential, and reduced survival, that were reminiscent of the traits observed in est2 budding yeast mutants. Telomerase activity was observed in wild-type fungus sporidia but not those of the disruption mutant. The introduction of a self-replicating plasmid expressing Trt1 into the mutant strain restored growth proficiency and replicative potential. Analyses of trt1 crosses in planta suggested that Trt1 is necessary for teliospore formation in homozygous disrupted diploids and that telomerase is haploinsufficient in heterozygous diploids. Additionally, terminal restriction fragment analysis in the progeny hinted at alternative survival mechanisms similar to those of budding yeast.
在本研究中,我们对双态真菌玉米黑粉菌中的端粒酶逆转录酶亚基进行了研究。该蛋白(Trt1)含有1371个氨基酸以及所有典型的TERT基序。通过破坏trt1产生的突变体具有衰老特征,如生长延迟、复制潜力低和存活率降低,这让人联想到在est2芽殖酵母突变体中观察到的特征。在野生型真菌担孢子中观察到了端粒酶活性,而破坏突变体的担孢子中未观察到。将表达Trt1的自我复制质粒导入突变菌株可恢复生长能力和复制潜力。对植物中trt1杂交的分析表明,Trt1对于纯合破坏二倍体中冬孢子的形成是必需的,并且端粒酶在杂合二倍体中是单倍体不足的。此外,后代中的末端限制片段分析暗示了类似于芽殖酵母的替代存活机制。