Borodulina Olga R, Kosushkin Sergey A, Ustyantsev Ilia G, Vassetzky Nikita S, Kramerov Dmitri A
Laboratory of Eukaryotic Genome Evolution, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, Russia.
Independent Researcher, Moscow 119991, Russia.
Noncoding RNA. 2025 May 14;11(3):39. doi: 10.3390/ncrna11030039.
SINEs (short interspersed elements) are eukaryotic non-autonomous retrotransposons. They are transcribed by RNA polymerase III (pol III) and generate non-coding RNAs. The 3' end of many mammalian SINEs contains a polyadenylation signal (AATAAA), a pol III transcription terminator, and an A-rich tail. Studies have shown that, in human HeLa cells that have been transiently transfected with such SINEs, short pol III-generated SINE transcripts undergo polyadenylation, resulting in the addition of a long poly(A)-tail. Notably, this AAUAAA-dependent polyadenylation is not characteristic of any other transcripts synthesized by pol III. B2 SINEs, found in the genomes of mouse-like rodents, exemplify all these features. In this study, we implemented a novel approach to sequencing pol III-generated B2 transcripts from mouse cell cultures (L929 and 4T1) and organs (brain and testis). Transcription occurred in 16,000-20,000 B2 copies in each cell type, 51-62% of which were transcribed in all four cell types. Effective transcription terminators (e.g., TCT and T) were found in approximately 40% of the transcribed B2 copies. The transcripts of these B2 copies contained a truncated terminator sequence, as pol III transcriptional arrest is known to occur within the terminator, with a poly(A)-tail immediately downstream. Such a tail could only have formed through RNA polyadenylation. These results demonstrate that B2 transcripts synthesized by pol III are capable of polyadenylation in mouse cells. We discuss the transcription of B2 copies with and without moderately efficient pol III terminators (TCTTT) and provide examples of the polyadenylation of such transcripts.
短散在元件(SINEs)是真核生物中的非自主逆转座子。它们由RNA聚合酶III(pol III)转录并产生非编码RNA。许多哺乳动物SINEs的3'端包含一个聚腺苷酸化信号(AATAAA)、一个pol III转录终止子和一个富含A的尾巴。研究表明,在瞬时转染了此类SINEs的人类HeLa细胞中,pol III产生的短SINE转录本会进行聚腺苷酸化,从而导致添加一个长的聚(A)尾巴。值得注意的是,这种依赖AAUAAA的聚腺苷酸化并非pol III合成的任何其他转录本的特征。在鼠类啮齿动物基因组中发现的B2 SINEs体现了所有这些特征。在本研究中,我们采用了一种新方法,对来自小鼠细胞培养物(L929和4T1)及器官(脑和睾丸)中pol III产生的B2转录本进行测序。在每种细胞类型中,16,000 - 20,000个B2拷贝发生转录,其中51 - 62%在所有四种细胞类型中都有转录。在大约40%的转录B2拷贝中发现了有效的转录终止子(如TCT和T)。这些B2拷贝的转录本包含一个截短的终止子序列,因为已知pol III转录终止发生在终止子内,紧接着下游是一个聚(A)尾巴。这样的尾巴只能通过RNA聚腺苷酸化形成。这些结果表明,pol III合成的B2转录本在小鼠细胞中能够进行聚腺苷酸化。我们讨论了带有和不带有中等效率pol III终止子(TCTTT)的B2拷贝的转录情况,并提供了此类转录本聚腺苷酸化的实例。