Charting DENR-dependent translation reinitiation uncovers predictive uORF features and links to circadian timekeeping via Clock
NUCLEIC ACIDS RESEARCH
Authors: Castelo-Szekely, Violeta; De Matos, Mara; Tusup, Marina; Pascolo, Steve; Ule, Jernej; Gatfield, David
Abstract
The non-canonical initiation factor DENR promotes translation reinitiation on mRNAs harbouring upstream open reading frames (uORFs). Moreover, DENR depletion shortens circadian period in mouse fibroblasts, suggesting involvement of uORF usage and reinitiation in clock regulation. To identify DENR-regulated translation events transcriptome-wide and, in particular, specific core clock transcripts affected by this mechanism, we have used ribosome profiling in DENR-deficient NIH3T3 cells. We uncovered 240 transcripts with altered translation rate, and used linear regression analysis to extract 5 UTR features predictive of DENR dependence. Among core clock genes, we identified Clock as a DENR target. Using Clock 5 UTR mutants, we mapped the specific uORF through which DENR acts to regulate CLOCK protein biosynthesis. Notably, these experiments revealed an alternative downstream start codon, likely representing the bona fide CLOCK N-terminus. Our findings provide insights into uORF-mediated translational regulation that can regulate the mammalian circadian clock and gene expression at large.
Using remote sensing to calculate plant available nitrogen needed by crops on swine factory farm sprayfields in North Carolina
REMOTE SENSING FOR AGRICULTURE, ECOSYSTEMS, AND HYDROLOGY XVII
Authors: Christenson, Elizabeth; Serre, Marc
Abstract
North Carolina (NC) is the second largest producer of hogs in the United States with Duplin county, NC having the densest population of hogs in the world. In NC, liquid swine manure is generally stored in open-air lagoons and sprayed onto sprayfields with sprinkler systems to be used as fertilizer for crops. Swine factory farms, termed concentrated animal feeding operations (CAFOs), are regulated by the Department of Environment and Natural Resources (DENR) based on nutrient management plans (NMPs) having balanced plant available nitrogen (PAN). The estimated PAN in liquid manure being sprayed must be less than the estimated PAN needed crops during irrigation. Estimates for PAN needed by crops are dependent on crop and soil types. Objectives of this research were to develop a new, time-efficient method to identify PAN needed by crops on Duplin county sprayfields for years 2010-2014. Using remote sensing data instead of NMP data to identify PAN needed by crops allowed calendar year identification of which crops were grown on sprayfields instead of a five-year range of values. Although permitted data have more detailed crop information than remotely sensed data, identification of PAN needed by crops using remotely sensed data is more time efficient, internally consistent, easily publically accessible, and has the ability to identify annual changes in PAN on sprayfields. Once PAN needed by crops is known, remote sensing can be used to quantify PAN at other spatial scales, such as sub-watershed levels, and can be used to inform targeted water quality monitoring of swine CAFOs.