ABSTRACT
SRSF1 governs splicing of over 1500 mRNA transcripts. SRSF1 contains two RNA-recognition motifs (RRMs) and a C-terminal Arg/Ser-rich region (RS). It has been thought that SRSF1 RRMs exclusively recognize single-stranded exonic splicing enhancers, while RS lacks RNA-binding specificity. With our success in solving the insolubility problem of SRSF1, we can explore the unknown RNA-binding landscape of SRSF1. We find that SRSF1 RS prefers purine over pyrimidine. Moreover, SRSF1 binds to the G-quadruplex (GQ) from the ARPC2 mRNA, with both RRMs and RS being crucial. Our binding assays show that the traditional RNA-binding sites on the RRM tandem and the Arg in RS are responsible for GQ binding. Interestingly, our FRET and circular dichroism data reveal that SRSF1 unfolds the ARPC2 GQ, with RS leading unfolding and RRMs aiding. Our saturation transfer difference NMR results discover that Arg residues in SRSF1 RS interact with the guanine base but not other nucleobases, underscoring the uniqueness of the Arg/guanine interaction. Our luciferase assays confirm that SRSF1 can alleviate the inhibitory effect of GQ on gene expression in the cell. Given the prevalence of RNA GQ and SR proteins, our findings unveil unexplored SR protein functions with broad implications in RNA splicing and translation.
Subject(s)
G-Quadruplexes , Protein Binding , Serine-Arginine Splicing Factors , Serine-Arginine Splicing Factors/metabolism , Serine-Arginine Splicing Factors/genetics , Serine-Arginine Splicing Factors/chemistry , Humans , Binding Sites , RNA Splicing , RNA Recognition Motif/genetics , RNA, Messenger/metabolism , RNA, Messenger/genetics , RNA, Messenger/chemistry , RNA/metabolism , RNA/genetics , RNA/chemistryABSTRACT
SRSF1 governs splicing of over 1,500 mRNA transcripts. SRSF1 contains two RNA-recognition motifs (RRMs) and a C-terminal Arg/Ser-rich region (RS). It has been thought that SRSF1 RRMs exclusively recognize single-stranded exonic splicing enhancers, while RS lacks RNA-binding specificity. With our success in solving the insolubility problem of SRSF1, we can explore the unknown RNA-binding landscape of SRSF1. We find that SRSF1 RS prefers purine over pyrimidine. Moreover, SRSF1 binds to the G-quadruplex (GQ) from the ARPC2 mRNA, with both RRMs and RS being crucial. Our binding assays show that the traditional RNA-binding sites on the RRM tandem and the Arg in RS are responsible for GQ binding. Interestingly, our FRET and circular dichroism data reveal that SRSF1 unfolds the ARPC2 GQ, with RS leading unfolding and RRMs aiding. Our saturation transfer difference NMR results discover that Arg residues in SRSF1 RS interact with the guanine base but other nucleobases, underscoring the uniqueness of the Arg/guanine interaction. Our luciferase assays confirm that SRSF1 can alleviate the inhibitory effect of GQ on gene expression in the cell. Given the prevalence of RNA GQ and SR proteins, our findings unveil unexplored SR protein functions with broad implications in RNA splicing and translation.
ABSTRACT
Efforts to reverse the obesity epidemic require policy, systems, and environmental (PSE) change strategies. Despite the availability of evidence-based and other promising PSE interventions, limited evidence exists on the "how-to" of transitioning them into practice. For the past 13 years, the Los Angeles County Department of Public Health has been building capacity among community residents and other stakeholders to create effective community coalitions and to implement well-designed policy strategy campaigns using an evidence-based approach to policy change, the policy adoption model (PAM). Implementing a phase-based approach to policy change, the PAM was initially used to support the passage of over 140 tobacco control and prevention policies in Los Angeles County. Following these successes, Los Angeles County Department of Public Health applied the PAM to obesity prevention, operationalizing the policy process by training community residents and other stakeholders on the use of the model. The PAM has shown to be helpful in promoting PSE change in tobacco control and obesity prevention, suggesting a local-level model potentially applicable to other fields of public health seeking sustainable, community-driven policy change.
