Mravinac Brankica, Meštrović Nevenka, Cavrak Vladimir Vanja, Plohl Miroslav
Division of Molecular Biology, Ruđer Bošković Institute, Bijenička, Zagreb, Croatia.
Chromosoma. 2011 Aug;120(4):367-76. doi: 10.1007/s00412-011-0317-x. Epub 2011 Apr 16.
The TTAGG repeat, the only determined telomerase-dependent sequence in the Insecta, is generally reputed to be the canonical telomeric motif within the class. By studying the distribution of telomeric DNAs in 30 coleopteran beetles using Southern hybridization, BAL 31 DNA end-degradation assay and fluorescence in situ hybridization, we showed that arrays built of a TCAGG repeat substitute for (TTAGG)n sequences in all tested species within the superfamily Tenebrionoidea. We also provided the experimental evidence that (TCAGG)n repeats represent the terminal sequences on all chromosomes of the model species Tribolium castaneum. (TCAGG)n repeats are therefore promoted as the first sequence-motif alternative to TTAGG-type chromosome ends in insects. Detection of species negative for both TTAGG and TCAGG reveals that, although widespread, these motifs are not ubiquitous telomeric sequences within the order Coleoptera. In addition, Timarcha balearica proved to be a species that harbors (TTAGG)n repeats, but not at telomeric positions, thus further increasing the complexity of telomeric DNAs. Our experiments discarded CTAGG, CTGGG, TTGGG, and TTAGGG variants as potential replacements in TTAGG/TCAGG-negative species, indicating that chromosome termini of these beetles comprise other form(s) of telomeric sequences and telomere maintenance mechanisms.
TTAGG重复序列是昆虫纲中唯一确定的依赖端粒酶的序列,通常被认为是该类群中的典型端粒基序。通过使用Southern杂交、BAL 31 DNA末端降解分析和荧光原位杂交研究30种鞘翅目甲虫中端粒DNA的分布,我们发现,在拟步甲总科的所有测试物种中,由TCAGG重复序列组成的阵列替代了(TTAGG)n序列。我们还提供了实验证据,证明(TCAGG)n重复序列代表模式物种赤拟谷盗所有染色体的末端序列。因此,(TCAGG)n重复序列被推举为昆虫中TTAGG型染色体末端的首个序列基序替代物。对TTAGG和TCAGG均呈阴性的物种的检测表明,尽管这些基序分布广泛,但它们并非鞘翅目内普遍存在的端粒序列。此外,巴利阿里岛Timarcha属甲虫被证明是一个含有(TTAGG)n重复序列但不在端粒位置的物种,从而进一步增加了端粒DNA的复杂性。我们的实验排除了CTAGG、CTGGG、TTGGG和TTAGGG变体作为TTAGG/TCAGG阴性物种的潜在替代物,这表明这些甲虫的染色体末端包含其他形式的端粒序列和端粒维持机制。