TY - JOUR T1 - Genome-scale reconstruction of Gcn4/ATF4 networks driving a growth program. JF - PLoS Genet Y1 - 2020 A1 - Srinivasan, Rajalakshmi A1 - Walvekar, Adhish S A1 - Rashida, Zeenat A1 - Seshasayee, Aswin A1 - Laxman, Sunil KW - Basic-Leucine Zipper Transcription Factors KW - Cell Proliferation KW - Gene Expression Regulation, Fungal KW - Gene Regulatory Networks KW - Genome, Fungal KW - Ribosomes KW - Saccharomyces cerevisiae KW - Saccharomyces cerevisiae Proteins KW - Transcriptional Activation AB -

Growth and starvation are considered opposite ends of a spectrum. To sustain growth, cells use coordinated gene expression programs and manage biomolecule supply in order to match the demands of metabolism and translation. Global growth programs complement increased ribosomal biogenesis with sufficient carbon metabolism, amino acid and nucleotide biosynthesis. How these resources are collectively managed is a fundamental question. The role of the Gcn4/ATF4 transcription factor has been best studied in contexts where cells encounter amino acid starvation. However, high Gcn4 activity has been observed in contexts of rapid cell proliferation, and the roles of Gcn4 in such growth contexts are unclear. Here, using a methionine-induced growth program in yeast, we show that Gcn4/ATF4 is the fulcrum that maintains metabolic supply in order to sustain translation outputs. By integrating matched transcriptome and ChIP-Seq analysis, we decipher genome-wide direct and indirect roles for Gcn4 in this growth program. Genes that enable metabolic precursor biosynthesis indispensably require Gcn4; contrastingly ribosomal genes are partly repressed by Gcn4. Gcn4 directly binds promoter-regions and transcribes a subset of metabolic genes, particularly driving lysine and arginine biosynthesis. Gcn4 also globally represses lysine and arginine enriched transcripts, which include genes encoding the translation machinery. The Gcn4 dependent lysine and arginine supply thereby maintains the synthesis of the translation machinery. This is required to maintain translation capacity. Gcn4 consequently enables metabolic-precursor supply to bolster protein synthesis, and drive a growth program. Thus, we illustrate how growth and starvation outcomes are both controlled using the same Gcn4 transcriptional outputs that function in distinct contexts.

VL - 16 IS - 12 ER - TY - JOUR T1 - Genome-Wide Analysis of Polyadenylation Events in Schmidtea mediterranea. JF - G3 (Bethesda) Y1 - 2016 A1 - Lakshmanan, Vairavan A1 - Bansal, Dhiru A1 - Kulkarni, Jahnavi A1 - Poduval, Deepak A1 - Krishna, Srikar A1 - Sasidharan, Vidyanand A1 - Anand, Praveen A1 - Seshasayee, Aswin A1 - Palakodeti, Dasaradhi KW - 3' Untranslated Regions KW - Animals KW - Computational Biology KW - Genome, Helminth KW - Genome-Wide Association Study KW - High-Throughput Nucleotide Sequencing KW - MicroRNAs KW - Molecular Sequence Annotation KW - Platyhelminths KW - Poly A KW - Polyadenylation KW - Reproducibility of Results KW - RNA Interference KW - RNA Processing, Post-Transcriptional KW - RNA, Messenger AB -

In eukaryotes, 3' untranslated regions (UTRs) play important roles in regulating posttranscriptional gene expression. The 3'UTR is defined by regulated cleavage/polyadenylation of the pre-mRNA. The advent of next-generation sequencing technology has now enabled us to identify these events on a genome-wide scale. In this study, we used poly(A)-position profiling by sequencing (3P-Seq) to capture all poly(A) sites across the genome of the freshwater planarian, Schmidtea mediterranea, an ideal model system for exploring the process of regeneration and stem cell function. We identified the 3'UTRs for ∼14,000 transcripts and thus improved the existing gene annotations. We found 97 transcripts, which are polyadenylated within an internal exon, resulting in the shrinking of the ORF and loss of a predicted protein domain. Around 40% of the transcripts in planaria were alternatively polyadenylated (ApA), resulting either in an altered 3'UTR or a change in coding sequence. We identified specific ApA transcript isoforms that were subjected to miRNA mediated gene regulation using degradome sequencing. In this study, we also confirmed a tissue-specific expression pattern for alternate polyadenylated transcripts. The insights from this study highlight the potential role of ApA in regulating the gene expression essential for planarian regeneration.

VL - 6 IS - 10 ER -