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• W3005070630 endingPage "982" @default.
• W3005070630 startingPage "970" @default.
• W3005070630 abstract "Cleavage and polyadenylation specificity factor 30 (CPSF30) is a zinc finger protein that plays a crucial role in the transition of pre-mRNA to RNA. CPSF30 contains five conserved CCCH domains and a CCHC zinc knuckle domain. CPSF30 activity is critical for pre-mRNA processing. A truncated form of the protein, in which only the CCCH domains are present, has been shown to specifically bind AU-rich pre-mRNA targets; however, the RNA binding and recognition properties of full-length CPSF30 are not known. Herein, we report the isolation and biochemical characterization of full-length CPSF30. We report that CPSF30 contains one 2Fe-2S cluster in addition to five zinc ions, as measured by inductively coupled plasma mass spectrometry, ultraviolet-visible spectroscopy, and X-ray absorption spectroscopy. Utilizing fluorescence anisotropy RNA binding assays, we show that full-length CPSF30 has high binding affinity for two types of pre-mRNA targets, AAUAAA and polyU, both of which are conserved sequence motifs present in the majority of pre-mRNAs. Binding to the AAUAAA motif requires that the five CCCH domains of CPSF30 be present, whereas binding to polyU sequences requires the entire, full-length CPSF30. These findings implicate the CCHC zinc knuckle present in the full-length protein as being critical for mediating polyU binding. We also report that truncated forms of the protein, containing either just two CCCH domains (ZF2 and ZF3) or the CCHC zinc knuckle domain, do not exhibit any RNA binding, indicating that CPSF30/RNA binding requires several ZF (and/or Fe-S cluster) domains working in concert to mediate RNA recognition." @default.
• W3005070630 created "2020-02-14" @default.
• W3005070630 creator A5014329266 @default.
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• W3005070630 date "2020-02-06" @default.
• W3005070630 modified "2023-09-24" @default.
• W3005070630 title "Unraveling the RNA Binding Properties of the Iron–Sulfur Zinc Finger Protein CPSF30" @default.
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• W3005070630 doi "https://doi.org/10.1021/acs.biochem.9b01065" @default.
• W3005070630 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/8575264" @default.
• W3005070630 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/32027124" @default.
• W3005070630 hasPublicationYear "2020" @default.
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