1. Structural basis of intron selection by U2 snRNP in the presence of covalent inhibitors
Patricia Gee, Vladimir Pena, Nicholas A Larsen, Andrew Cook, Anant Agrawal, Xiang Liu, Arun K Ghosh, Tuong-Vi Nguyen, Constantin Cretu, Melissa Jurica Nat Commun . 2021 Jul 23;12(1):4491. doi: 10.1038/s41467-021-24741-1.
Intron selection during the formation of prespliceosomes is a critical event in pre-mRNA splicing. Chemical modulation of intron selection has emerged as a route for cancer therapy. Splicing modulators alter the splicing patterns in cells by binding to the U2 snRNP (small nuclear ribonucleoprotein)-a complex chaperoning the selection of branch and 3' splice sites. Here we report crystal structures of the SF3B module of the U2 snRNP in complex with spliceostatin and sudemycin FR901464 analogs, and the cryo-electron microscopy structure of a cross-exon prespliceosome-like complex arrested with spliceostatin A. The structures reveal how modulators inactivate the branch site in a sequence-dependent manner and stall an E-to-A prespliceosome intermediate by covalent coupling to a nucleophilic zinc finger belonging to the SF3B subunit PHF5A. These findings support a mechanism of intron recognition by the U2 snRNP as a toehold-mediated strand invasion and advance an unanticipated drug targeting concept.
2. Enantioselective syntheses of FR901464 and spliceostatin A: potent inhibitors of spliceosome
Zhi-Hua Chen, Arun K Ghosh Org Lett . 2013 Oct 4;15(19):5088-91. doi: 10.1021/ol4024634.
Enantioselective syntheses of FR901464 and spliceostatin A, potent spliceosome inhibitors, are described. The synthesis of FR901464 has been accomplished in a convergent manner in 10 linear steps (20 total steps). The A-tetrahydropyran ring was constructed from (R)-isopropylidene glyceraldehyde. The functionalized tetrahydropyran B-ring was synthesized utilizing a Corey-Bakshi-Shibata reduction, an Achmatowicz reaction, and a stereoselective Michael addition as the key steps. Coupling of A- and B-ring fragments was accomplished via cross-metathesis.
3. Cytotoxic Spliceostatin Analogs from Pseudomonas sp
Lie-Feng Ma, Hong Zhao, Lin-Mei Shi, Zha-Jun Zhan, Bei Zhang Chem Biodivers . 2019 Sep;16(9):e1900266. doi: 10.1002/cbdv.201900266.
Two new spliceostatin analogs, designed as spliceostatins J and K (1 and 2), were isolated and identified from the culture of Pseudomonas sp., along with two known ones, FR901464 (3) and spliceostatin E (4). Their structures were elucidated by detailed interpretation of their spectroscopic data, especially 2D-NMR and HR-ESI-MS. Spliceostatin J (1) represented the first example of spliceostatins bearing an unusual hexahydrofuro[3,4-b]furan moiety. Biological assay showed all the isolated compounds except 1 displayed potent cytotoxic activities against two cancer cell lines (MDA-MB-231 and A-549). Structure-activity-relationship studies revealed that the tetrahydropyran ring in spliceostatin analogs was necessary for their bioactive retention.