Fine gene expression regulation by minor sequence variations downstream of the polyadenylation signal

Lidiya V. Boldyreva, Lyubov A. Yarinich, Elena N. Kozhevnikova, Anton V. Ivankin, Mikhail O. Lebedev, Alexey V. Pindyurin

Результат исследования: Научные публикации в периодических изданияхстатьярецензирование

1 Цитирования (Scopus)


The termination of transcription is a complex process that substantially contributes to gene regulation in eukaryotes. Previously, it was noted that a single cytosine deletion at the position + 32 bp relative to the single polyadenylation signal AAUAAA (hereafter the dC mutation) causes a 2-fold increase in the transcription level of the upstream eGFP reporter in mouse embryonic stem cells. Here, we analyzed the conservation of this phenomenon in immortalized mouse, human and drosophila cell lines and the influence of the dC mutation on the choice of the pre-mRNA cleavage sites. We have constructed dual-reporter plasmids to accurately measure the effect of the dC and other nearby located mutations on eGFP mRNA level by RT-qPCR. In this way, we found that the dC mutation leads to a 2-fold increase in the expression level of the upstream eGFP reporter gene in cultured mouse and human, but not in drosophila cells. In addition, 3′ RACE analysis demonstrated that eGFP pre-mRNAs are cut at multiple positions between + 14 to + 31, and that the most proximal cleavage site becomes almost exclusively utilized in the presence of the dC mutation. We also identified new short sequence variations located within positions + 25. + 40 and + 33. + 48 that increase eGFP expression up to ~2-4-fold. Altogether, the positive effect of the dC mutation seems to be conserved in mouse embryonic stem cells, mouse embryonic 3T3 fibroblasts and human HEK293T cells. In the latter cells, the dC mutation appears to be involved in regulating pre-mRNA cleavage site selection. Finally, a multiplexed approach is proposed to identify motifs located downstream of cleavage site(s) that are essential for transcription termination.

Язык оригиналаанглийский
Страницы (с-по)1539-1547
Число страниц9
ЖурналMolecular Biology Reports
Номер выпуска2
СостояниеОпубликовано - фев 2021


Подробные сведения о темах исследования «Fine gene expression regulation by minor sequence variations downstream of the polyadenylation signal». Вместе они формируют уникальный семантический отпечаток (fingerprint).