The Polyadenosine RNA-binding Protein, Zinc Finger Cys(3)His Protein 14 (ZC3H14), Regulates the Pre-mRNA Processing of a Key ATP Synthase Subunit mRNA

• Type of Material: Article
• Authors: Callie P. Wigington, Emory University; Kevin J. Morris, Emory University; Laura E. Newman, Emory University; Anita Corbett, Emory University
• Language: English
• Date: 2016-10-21
• Publisher: American Society for Biochemistry and Molecular Biology
• Publication Version: Final Published Version
• Copyright Statement: © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.
• Final Published Version (URL): http://dx.doi.org/10.1074/jbc.M116.754069
• Title of Journal or Parent Work: Journal of Biological Chemistry
• ISSN: 0021-9258
• Volume: 291
• Issue: 43
• Start Page: 22442
• End Page: 22459
• Grant/Funding Information: The Emory Neuroscience Microscopy Core facilities was a recipient of National Institutes of Health Grant P30NS055077.; This work was supported by the National Institutes of Health Grants R01GM058728 (to A. H. C.), R01GM090158 (to R. A. Kahn in support of L. N.), F31CA168321 (to C. P.W.), and T32GM008367 (to C. P.W. and L. N.) and by American Heart Association Grant 14PRE18840040 (to L. N.).
• Keywords: NUCLEAR POLY(A)-BINDING PROTEIN; RNA splicing; Biochemistry & Molecular Biology; RNA; Nab2; APOPTOSIS; post-transcriptional regulation; SACCHAROMYCES-CEREVISIAE; CELL VIABILITY; MSUT2; ZC3H14; ATP synthase; SPECIFICITY FACTOR; GENE-EXPRESSION; RNA processing; Science & Technology; MITOCHONDRIAL MORPHOLOGY; POLY(A) TAIL LENGTH; Life Sciences & Biomedicine; RNA-binding protein; CYTOCHROME-C; BROWN ADIPOSE-TISSUE
• Research Categories: Chemistry, Biochemistry; Biology, Cell

Abstract

Polyadenosine RNA-binding proteins (Pabs) regulate multiple steps in gene expression. This protein family includes the well studied Pabs, PABPN1 and PABPC1, as well as the newly characterized Pab, zinc finger CCCH-type containing protein 14 (ZC3H14). Mutations in ZC3H14 are linked to a form of intellectual disability. To probe the function of ZC3H14, we performed a transcriptome-wide analysis of cells depleted of either ZC3H14 or the control Pab, PABPN1. Depletion of PABPN1 affected - 17% of expressed transcripts, whereas ZC3H14 affected only - 1% of expressed transcripts. To assess the function of ZC3H14 in modulating target mRNAs, we selected the gene encoding the ATP synthase F 0 subunit C (ATP5G1) transcript. Knockdown of ZC3H14 significantly reduced ATP5G1 steady-state mRNA levels. Consistent with results suggesting that ATP5G1 turnover increases upon depletion of ZC3H14, double knockdown of ZC3H14 and the nonsense-mediated decay factor, UPF1, rescues ATP5G1 transcript levels. Furthermore, fractionation reveals an increase in the amount of ATP5G1 pre-mRNA that reaches the cytoplasm when ZC3H14 is depleted and that ZC3H14 binds to ATP5G1 pre-mRNA in the nucleus. These data support a role for ZC3H14 in ensuring proper nuclear processing and retention of ATP5G1 pre-mRNA. Consistent with the observation that ATP5G1 is a rate-limiting component for ATP synthase activity, knockdown of ZC3H14 decreases cellular ATP levels and causes mitochondrial fragmentation. These data suggest that ZC3H14 modulates pre-mRNA processing of select mRNA transcripts and plays a critical role in regulating cellular energy levels, observations that have broad implications for proper neuronal function.