Global analysis of pre-mRNA subcellular localization following splicing inhibition by spliceostatin A
- PMID: 27754875
- PMCID: PMC5159648
- DOI: 10.1261/rna.058065.116
Global analysis of pre-mRNA subcellular localization following splicing inhibition by spliceostatin A
Abstract
Spliceostatin A (SSA) is a methyl ketal derivative of FR901464, a potent antitumor compound isolated from a culture broth of Pseudomonas sp no. 2663. These compounds selectively bind to the essential spliceosome component SF3b, a subcomplex of the U2 snRNP, to inhibit pre-mRNA splicing. However, the mechanism of SSA's antitumor activity is unknown. It is noteworthy that SSA causes accumulation of a truncated form of the CDK inhibitor protein p27 translated from CDKN1B pre-mRNA, which is involved in SSA-induced cell-cycle arrest. However, it is still unclear whether pre-mRNAs are uniformly exported from the nucleus following SSA treatment. We performed RNA-seq analysis on nuclear and cytoplasmic fractions of SSA-treated cells. Our statistical analyses showed that intron retention is the major consequence of SSA treatment, and a small number of intron-containing pre-mRNAs leak into the cytoplasm. Using a series of reporter plasmids to investigate the roles of intronic sequences in the pre-mRNA leakage, we showed that the strength of the 5' splice site affects pre-mRNA leakage. Additionally, we found that the level of pre-mRNA leakage is related to transcript length. These results suggest that the strength of the 5' splice site and the length of the transcripts are determinants of the pre-mRNA leakage induced by SF3b inhibitors.
Keywords: RNA-seq; pre-mRNA nuclear retention; pre-mRNA splicing; spliceostatin A.
© 2016 Yoshimoto et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.
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References
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- Berglund JA, Chua K, Abovich N, Reed R, Rosbash M. 1997. The splicing factor BBP interacts specifically with the pre-mRNA branchpoint sequence UACUAAC. Cell 89: 781–787. - PubMed
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