|
ИСТИНА |
Войти в систему Регистрация |
ИПМех РАН |
||
The expression of a nested alternative open reading frame may be a novel mechanism for intronless gene expression controlтезисы доклада
- Авторы: Komarova T.V., Sheshukova E.V., Ershova N.M., Dorokhov Y.L.
- Сборник: The 43rd FEBS Open Bio:
- Серия: Suppl 1
- Том: 8
- Тезисы
- Год издания: 2018
- Место издания: FEBS Press Prague
- Первая страница: 114
- Последняя страница: 115
- DOI: 10.1002/2211-5463.12453
- Аннотация: The regulation of gene expression in animal and plant tissues at different stages of development and after exposure to external stimuli occurs at the transcriptional, post-transcriptional and post-translational levels. Recently, in the Solanaceae family of plants, we found a gene for the Kunitz protease inhibitor-like protein (KPILP) with low mRNA levels in the leaves of an intact plant (unlike what is observed in the roots). However, these levels sharply increased after a 72-h incubation in the dark under biotic stresses. Unlike the KPI in animals, the KPILP of Solanaceae plants does not contain introns, which excludes the regulation of its expression by a mechanism of alternative splicing. This phenomenon is characteristic of mammalian KPIs. To identify the mechanism for the regulation of Solanaceae KPILPs, we identified and examined the function of the KPILP nested alternative open reading frame (aORF) that controls the expression of maternal mRNA. Here, we investigated the regulation of KPILP under stress conditions at the level of gene transcription. To this end, we isolated a portion of Nicotiana benthamiana chromosomal DNA upstream of the KPILP coding sequence (proKPILP). We found that the segment directed the expression of the E. coli UidA reporter gene that encoded the β-D-glucuronidase (GUS) in plant agroinoculation experiments. A bioinformatic analysis revealed potential light-responsive elements in the proKPILP sequence. To understand if these elements influence proKPILP activity, agroinjected plants were isolated for 3 days in complete darkness. Then, the level of GUS and corresponding mRNA content were assessed in the leaves and compared with the constitutive CaMV 35S promoter. The results showed that, as in the case of the CaMV 35S promoter, darkness had a negligible effect on the activity of the proKPILP. We concluded that the stress-dependent content of the KPILP mRNA in the leaves is determined mainly by the aORF-mediated mechanism of maternal mRNA control. Thus, Solanaceae KPILP is an intronless matryoshka gene and is the first example of a gene expression regulation mechanism that is probably characteristic for intronless genes. This study was performed with financial support from the Russian Foundation for Basic Research (project No. 17-29-08012).
- Добавил в систему: Дорохов Юрий Леонидович