TY - JOUR T1 - Small Genetic Circuits and MicroRNAs: Big Players in Polymerase II Transcriptional Control in Plants. JF - Plant Cell Y1 - 2016 A1 - Megraw, Molly A1 - Cumbie, Jason S A1 - Ivanchenko, Maria G A1 - Filichkin, Sergei A AB -

RNA Polymerase II (Pol II) regulatory cascades involving transcription factors (TFs) and their targets orchestrate the genetic circuitry of every eukaryotic organism. In order to understand how these cascades function, they can be dissected into small genetic networks, each containing just a few Pol II transcribed genes, that generate specific signal-processing outcomes. Small RNA regulatory circuits involve direct regulation of a small RNA by a TF and/or direct regulation of a TF by a small RNA and have been shown to play unique roles in many organisms. Here, we will focus on small RNA regulatory circuits containing Pol II transcribed microRNAs (miRNAs). While the role of miRNA-containing regulatory circuits as modular building blocks for the function of complex networks has long been on the forefront of studies in the animal kingdom, plant studies are poised to take a lead role in this area because of their advantages in probing transcriptional and posttranscriptional control of Pol II genes. The relative simplicity of tissue- and cell-type organization, miRNA targeting, and genomic structure make the Arabidopsis thaliana plant model uniquely amenable for small RNA regulatory circuit studies in a multicellular organism. In this Review, we cover analysis, tools, and validation methods for probing the component interactions in miRNA-containing regulatory circuits. We then review the important roles that plant miRNAs are playing in these circuits and summarize methods for the identification of small genetic circuits that strongly influence plant function. We conclude by noting areas of opportunity where new plant studies are imminently needed.

VL - 28 IS - 2 ER - TY - JOUR T1 - Sustained-input switches for transcription factors and microRNAs are central building blocks of eukaryotic gene circuits. JF - Genome Biol Y1 - 2013 A1 - Megraw, Molly A1 - Mukherjee, Sayan A1 - Ohler, Uwe KW - Algorithms KW - Animals KW - Arabidopsis KW - Computational Biology KW - Drosophila melanogaster KW - Gene Expression Regulation KW - Gene Regulatory Networks KW - Humans KW - MicroRNAs KW - Molecular Sequence Annotation KW - Nucleic Acid Conformation KW - Software KW - Transcription Factors AB -

WaRSwap is a randomization algorithm that for the first time provides a practical network motif discovery method for large multi-layer networks, for example those that include transcription factors, microRNAs, and non-regulatory protein coding genes. The algorithm is applicable to systems with tens of thousands of genes, while accounting for critical aspects of biological networks, including self-loops, large hubs, and target rearrangements. We validate WaRSwap on a newly inferred regulatory network from Arabidopsis thaliana, and compare outcomes on published Drosophila and human networks. Specifically, sustained input switches are among the few over-represented circuits across this diverse set of eukaryotes.

VL - 14 IS - 8 ER - TY - JOUR T1 - A stele-enriched gene regulatory network in the Arabidopsis root. JF - Mol Syst Biol Y1 - 2011 A1 - Brady, Siobhan M A1 - Zhang, Lifang A1 - Megraw, Molly A1 - Martinez, Natalia J A1 - Jiang, Eric A1 - Yi, Charles S A1 - Liu, Weilin A1 - Zeng, Anna A1 - Taylor-Teeples, Mallorie A1 - Kim, Dahae A1 - Ahnert, Sebastian A1 - Ohler, Uwe A1 - Ware, Doreen A1 - Walhout, Albertha J M A1 - Benfey, Philip N KW - Arabidopsis KW - Arabidopsis Proteins KW - Gene Expression Profiling KW - Gene Regulatory Networks KW - MicroRNAs KW - Plant Roots KW - Reproducibility of Results KW - Systems Biology KW - Transcription Factors KW - Two-Hybrid System Techniques AB -

Tightly controlled gene expression is a hallmark of multicellular development and is accomplished by transcription factors (TFs) and microRNAs (miRNAs). Although many studies have focused on identifying downstream targets of these molecules, less is known about the factors that regulate their differential expression. We used data from high spatial resolution gene expression experiments and yeast one-hybrid (Y1H) and two-hybrid (Y2H) assays to delineate a subset of interactions occurring within a gene regulatory network (GRN) that determines tissue-specific TF and miRNA expression in plants. We find that upstream TFs are expressed in more diverse cell types than their targets and that promoters that are bound by a relatively large number of TFs correspond to key developmental regulators. The regulatory consequence of many TFs for their target was experimentally determined using genetic analysis. Remarkably, molecular phenotypes were identified for 65% of the TFs, but morphological phenotypes were associated with only 16%. This indicates that the GRN is robust, and that gene expression changes may be canalized or buffered.

VL - 7 ER -