Availability of splicing factors in the nucleoplasm can regulate the release of mRNA from the gene after transcription

Autoři: Hodaya Hochberg-Laufer aff001;  Noa Neufeld aff001;  Yehuda Brody aff001;  Shani Nadav-Eliyahu aff001;  Rakefet Ben-Yishay aff001;  Yaron Shav-Tal aff001
Působiště autorů: The Mina & Everard Goodman Faculty of Life Sciences & Institute of Nanotechnology, Bar-Ilan University, Ramat Gan, Israel aff001
Vyšlo v časopise: Availability of splicing factors in the nucleoplasm can regulate the release of mRNA from the gene after transcription. PLoS Genet 15(11): e32767. doi:10.1371/journal.pgen.1008459
Kategorie: Research Article
doi: 10.1371/journal.pgen.1008459


Gene expression dynamics can be measured in single living cells. Using a detectable transcriptionally active gene in living cells, we previously found that an mRNA undergoing several splicing events was retained at this gene after transcription until completion of mRNA processing. To determine the reason for this delay in release and whether mRNA retention on the gene might depend on splicing factor availability, we modulated the levels of splicing factors in the nucleus. Increasing the abundance of the diffusing fraction of splicing factors by their overexpression or by Clk1 kinase overexpression to disassemble nuclear speckles, led to a reduction in splicing factor residence times on the active gene, and the retained mRNA was rapidly released from the gene. Other treatments such as overexpression of a mutant inactive Clk1, the downregulation of MALAT1 lncRNA or of the Son protein, or the overexpression of the splicing factor import factor TNPO3, did not affect the dynamics of mRNA release from the gene. We found that the faster release of the mRNA from the gene mediated by increased availability of splicing factors, was dependent on the RS domain of the splicing factors and its phosphorylation state. We propose that the relative abundancies of splicing factors in the nucleoplasm can affect their availability for the splicing events taking place, and regulate the kinetics of mRNA release from the gene after processing.

Klíčová slova:

DNA transcription – Hyperexpression techniques – Introns – Messenger RNA – Phosphorylation – RNA splicing – Small interfering RNAs – Fluorescence recovery after photobleaching


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PLOS Genetics

2019 Číslo 11

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