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Published on: March 4, 2014
Determination of trunk neural crest cell fate and susceptibility to splicing perturbation by the DLC1-SF3B1-PHF5A
Zhengfan Zheng1, Suisui Guo1,2, Hoi Yau Tam1
1School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
Abstract:
How the ubiquitously expressed splicing factors specifically regulate neural crest (NC) development and enhance their vulnerability to splicing perturbations remain poorly understood. Here, we show that NC-specific DLC1, partnering with SF3B1-PHF5A splicing complex, are crucial for determining avian trunk NC cell fate by regulating the splicing of NC specifiers SOX9 and SNAI2 pre-mRNAs rather than their upstream regulators BMP4, WNT1, and PAX7. Mechanistically, SF3B1-PHF5A binds to the intronic branch site (BS) sequences of all factors, while DLC1 interacts with a specific motif near the BS sequences of SOX9 and SNAI2, thereby determining their functional specificity in NC specification. Moreover, DLC1 increases NC cells' vulnerability to splicing modulator pladienolide B (PB) by reducing the binding capacity of the SF3B1-PHF5A splicing complex to the shorter length of both SOX9 intron 2 and SNAI2 intron 1, which possess weaker polypyrimidine tract 3' of the BS sequence, resulting in intron retention and loss of NC progenitors. Conversely, somite specific SLU7-SF3B1-PHF5A splicing complex regulates SOX9 and SNAI2 expression and imparts resistance to PB. Our data reveal the cell-type specific splicing complexes with distinct vulnerabilities to PB, highlighting the critical role of the DLC1-SF3B1-PHF5A in determining trunk NC cell fate and enhancing its susceptibility to splicing perturbation.
Insights
Neural crest (NC) development relies on specific splicing factors like DLC1-SF3B1-PHF5A, which regulate key genes. This complex enhances NC cell vulnerability to splicing perturbations, impacting cell fate.
Area of Science:
- Developmental Biology
- Molecular Biology
- Genetics
Background:
- Splicing factors regulate gene expression but their specific roles in neural crest (NC) development are unclear.
- Understanding how these factors influence NC cell fate and vulnerability to splicing perturbations is crucial.
Purpose of the Study:
- To investigate the role of NC-specific splicing factors in avian trunk NC cell fate determination.
- To elucidate the mechanism by which DLC1-SF3B1-PHF5A complex regulates NC specifiers and confers vulnerability to splicing modulators.
Main Methods:
- Investigated the interaction of DLC1 with the SF3B1-PHF5A splicing complex.
- Analyzed the regulation of SOX9 and SNAI2 pre-mRNA splicing by NC-specific and somite-specific splicing complexes.
- Assessed the impact of splicing modulator pladienolide B (PB) on NC cell progenitors.
Main Results:
- NC-specific DLC1-SF3B1-PHF5A complex regulates SOX9 and SNAI2 splicing, determining trunk NC cell fate.
- DLC1 enhances NC cell vulnerability to PB by disrupting SF3B1-PHF5A binding to specific introns, leading to intron retention.
- Somite-specific SLU7-SF3B1-PHF5A complex confers resistance to PB.
Conclusions:
- Splicing complexes are cell-type specific and exhibit distinct vulnerabilities to splicing perturbations.
- The DLC1-SF3B1-PHF5A complex plays a critical role in trunk NC cell fate and increases susceptibility to splicing interference.
