@article {753, title = {Metabolomics analysis reveals both plant variety and choice of hormone treatment modulate vinca alkaloid production in Catharanthus roseus}, journal = {Plant Direct}, volume = {4}, year = {2020}, month = {09/2020}, abstract = {

The medicinal plant\ Catharanthus roseus\ produces numerous secondary metabolites of interest for the treatment of many diseases \– most notably for the terpene indole alkaloid (TIA) vinblastine, which is used in the treatment of leukemia and Hodgkin\&$\#$39;s lymphoma. Historically, methyl jasmonate (MeJA) has been used to induce TIA production, but in the past, this has only been investigated in whole seedlings, cell culture, or hairy root culture. This study examines the effects of the phytohormones MeJA and ethylene on the induction of TIA biosynthesis and accumulation in the shoots and roots of 8-day-old seedlings of two varieties of\ C. roseus. Using LCMS and RT-qPCR, we demonstrate the importance of variety selection, as we observe markedly different induction patterns of important TIA precursor compounds. Additionally, both phytohormone choice and concentration have significant effects on TIA biosynthesis. Finally, our study suggests that several early-induction pathway steps as well as pathway-specific genes are likely to be transcriptionally regulated. Our findings highlight the need for a complete set of\&$\#$39;omics resources in commonly used\ C. roseus\ varieties and the need for caution when extrapolating results from one cultivar to another.

}, url = {https://onlinelibrary.wiley.com/doi/10.1002/pld3.267}, author = {Valerie N. Fraser and Benjamin Philmus and Molly Megraw} } @article {750, title = {Arabidopsis bioinformatics resources: The current state, challenges, and priorities for the future}, journal = {Plant Direct}, volume = {3}, year = {2019}, month = {01/2019}, url = {https://onlinelibrary.wiley.com/doi/full/10.1002/pld3.109}, author = {Colleen Doherty and Joanna Friesner and Brian Gregory and Ann Loraine and Molly Megraw and Nicholas Provart and R Keith Slotkin and Chris Town and Sarah M Assmann and Michael Axtell and Tanya Berardini and Sixue Chen and Malia Gehan and Eva Huala and Pankaj Jaiswal and Stephen Larson and Song Li and Sean May and Todd Michael and Chris Pires and Chris Topp and Justin Walley and Eve Wurtele} } @article {746, title = {The Next Generation of Training for Arabidopsis Researchers: Bioinformatics and Quantitative Biology}, journal = {Plant Physiology}, volume = {175}, year = {2017}, month = {12/2017}, pages = {1499-1509}, abstract = {
It has been more than 50 years since Arabidopsis (Arabidopsis thaliana) was first introduced as a model organism to understand basic processes in plant biology. A well-organized scientific community has used this small reference plant species to make numerous fundamental plant biology discoveries (Provart et al., 2016). Due to an extremely well-annotated genome and advances in high-throughput sequencing, our understanding of this organism and other plant species has become even more intricate and complex. Computational resources, including CyVerse,3 Araport,4 The Arabidopsis Information Resource (TAIR),5 and BAR,6 have further facilitated novel findings with just the click of a mouse. As we move toward understanding biological systems, Arabidopsis researchers will need to use more quantitative and computational approaches to extract novel biological findings from these data. Here, we discuss guidelines, skill sets, and core competencies that should be considered when developing curricula or training undergraduate or graduate students, postdoctoral researchers, and faculty. A selected case study provides more specificity as to the concrete issues plant biologists face and how best to address such challenges.
}, url = {http://www.plantphysiol.org/content/175/4/1499}, author = {Joanna Friesner and Sarah M. Assmann and Ruth Bastow and Julia Bailey-Serres and Jim Beynon and Volker Brendel and C. Robin Buell and Alexander Bucksch and Wolfgang Busch and Taku Demura and Jose R. Dinneny and Colleen J. Doherty and Andrea L. Eveland and Pascal Falter-Braun and Malia A. Gehan and Michael Gonzales and Erich Grotewold and Rodrigo Gutierrez and Ute Kramer and Gabriel Krouk and Shisong Ma and R.J. Cody Markelz and Molly Megraw and Blake C. Meyers and James A.H. Murray and Nicholas J. Provart and Sue Rhee and Roger Smith and Edgar P. Spalding and Crispin Taylor and Tracy K. Teal and Keiko U. Torii and Chris Town and Matthew Vaughn and Richard Vierstra and Doreen Ware and Olivia Wilkins and Cranos Williams and Siobhan M. Brady} } @article {742, title = {Establishment of Expression in the SHORTROOT-SCARECROW Transcriptional Cascade through Opposing Activities of Both Activators and Repressors}, journal = {Dev Cell}, volume = {39}, year = {2016}, month = {12/2016}, pages = {585-596}, abstract = {

Tissue-specific gene expression is often thought to arise from spatially restricted transcriptional cascades. However, it is unclear how expression is established at the top of these cascades in the absence of pre-existing specificity. We generated a transcriptional network to explore how transcription factor expression is established in the Arabidopsis thaliana root ground tissue. Regulators of the SHORTROOT-SCARECROW transcriptional cascade were validated in planta. At the top of this cascade, we identified both activators and repressors of SHORTROOT. The aggregate spatial expression of these regulators is not sufficient to predict transcriptional specificity. Instead, modeling, transcriptional reporters, and synthetic promoters support a mechanism whereby expression at the top of the SHORTROOT-SCARECROW cascade is established through opposing activities of activators and repressors.

}, keywords = {Arabidopsis Proteins/ genetics/ metabolism, Arabidopsis/ genetics/growth \& development/ metabolism, Computer Simulation, Gene Expression Regulation, Plant, Gene Regulatory Networks, Genes, Plant, Genes, Reporter, Genes, Synthetic, Models, Genetic, Plant Roots/cytology/metabolism, Plants, Genetically Modified, Promoter Regions, Genetic, Repressor Proteins/genetics/metabolism, Trans-Activators/genetics/metabolism, Transcription Factors/ genetics/ metabolism, Two-Hybrid System Techniques}, isbn = {1878-1551 (Electronic)1534-5807 (Linking)}, url = {https://doi.org/10.1016/j.devcel.2016.09.031}, author = {Sparks, E. E. and Drapek, C. and Gaudinier, A. and Li, S. and Ansariola, M. and Shen, N. and Hennacy, J. H. and Zhang, J. and Turco, G. and Petricka, J. J. and Foret, J. and Hartemink, A. J. and Gordan, R. and Megraw, M. and Brady, S. M. and Benfey, P. N.} } @article {314, title = {Alternative splicing in plants: directing traffic at the crossroads of adaptation and environmental stress.}, journal = {Curr Opin Plant Biol}, volume = {24}, year = {2015}, month = {2015 Apr}, pages = {125-35}, abstract = {

In recent years, high-throughput sequencing-based analysis of plant transcriptomes has suggested that up to \∼60\% of plant gene loci encode alternatively spliced mature transcripts. These studies have also revealed that alternative splicing in plants can be regulated by cell type, developmental stage, the environment, and the circadian clock. Alternative splicing is coupled to RNA surveillance and processing mechanisms, including nonsense mediated decay. Recently, non-protein-coding transcripts have also been shown to undergo alternative splicing. These discoveries collectively describe a robust system of post-transcriptional regulatory feedback loops which influence RNA abundance. In this review, we summarize recent studies describing the specific roles alternative splicing and RNA surveillance play in plant adaptation to environmental stresses and the regulation of the circadian clock.

}, keywords = {Adaptation, Biological, Alternative Splicing, Circadian Clocks, Plant Physiological Phenomena, RNA, Plant, Stress, Physiological}, issn = {1879-0356}, doi = {10.1016/j.pbi.2015.02.008}, author = {Filichkin, Sergei and Priest, Henry D and Megraw, Molly and Mockler, Todd C} } @article {315, title = {Environmental stresses modulate abundance and timing of alternatively spliced circadian transcripts in Arabidopsis.}, journal = {Mol Plant}, volume = {8}, year = {2015}, month = {2015 Feb}, pages = {207-27}, abstract = {

Environmental stresses profoundly altered accumulation of nonsense mRNAs including intron-retaining (IR) transcripts in Arabidopsis. Temporal patterns of stress-induced IR mRNAs were dissected using both oscillating and non-oscillating transcripts. Broad-range thermal cycles triggered a sharp increase in the long IR CCA1 isoforms and altered their phasing to different times of day. Both abiotic and biotic stresses such as drought or Pseudomonas syringae infection induced a similar increase. Thermal stress induced a time delay in accumulation of CCA1 I4Rb transcripts, whereas functional mRNA showed steady oscillations. Our data favor a hypothesis that stress-induced instabilities of the central oscillator can be in part compensated through fluctuations in abundance and out-of-phase oscillations of CCA1 IR transcripts. Taken together, our results support a concept that mRNA abundance can be modulated through altering ratios between functional and nonsense/IR transcripts. SR45 protein specifically bound to the retained CCA1 intron in\ vitro, suggesting that this splicing factor could be involved in regulation of intron retention. Transcriptomes of nonsense-mediated mRNA decay (NMD)-impaired and heat-stressed plants shared a set of retained introns associated with stress- and defense-inducible transcripts. Constitutive activation of certain stress response networks in an NMD mutant could be linked to disequilibrium between functional and nonsense mRNAs.

}, keywords = {Alternative Splicing, Arabidopsis, Arabidopsis Proteins, Circadian Clocks, Gene Expression Regulation, Plant, Introns, Nonsense Mediated mRNA Decay}, issn = {1752-9867}, doi = {10.1016/j.molp.2014.10.011}, author = {Filichkin, Sergei A and Cumbie, Jason S and Dharmawardhana, Palitha and Jaiswal, Pankaj and Chang, Jeff H and Palusa, Saiprasad G and Reddy, A S N and Megraw, Molly and Mockler, Todd C} } @article {317, title = {Environmental Stresses Modulate Abundance and Timing of Alternatively Spliced Circadian Transcripts in Arabidopsis.}, journal = {Mol Plant}, year = {2014}, month = {2014 Nov 3}, abstract = {

Environmental stresses profoundly altered accumulation of nonsense mRNAs including intron retaining (IR) transcripts in Arabidopsis. Temporal patterns of stress-induced IR mRNAs were dissected using both oscillating and non-oscillating transcripts. Broad range thermal cycles triggered a sharp increase in the long intron retaining CCA1 isoforms and altered their phasing to different times of day. Both abiotic and biotic stresses such as drought or P. syringae infection induced similar increase. Thermal stress induced a time delay in accumulation of CCA1 I4Rb transcripts whereas functional mRNA showed steady oscillations. Our data favor a hypothesis that stress-induced instabilities of the central oscillator can be in part compensated through fluctuations in abundance and out of phase oscillations of CCA1 IR transcripts. Altogether, our results support a concept that mRNA abundance can be modulated through altering ratios between functional and nonsense/IR transcripts. SR45 protein specifically bound to the retained CCA1 intron in vitro, suggesting that this splicing factor could be involved in regulation of intron retention. Transcriptomes of NMD-impaired and heat-stressed plants shared a set of retained introns associated with stress- and defense-inducible transcripts. Constitutive activation of certain stress response networks in an NMD mutant could be linked to disequilibrium between functional and nonsense mRNAs.

}, issn = {1752-9867}, doi = {10.1093/mp/ssu130}, author = {Filichkin, Sergei A and Cumbie, Jason S and Dharmawadhana, J Palitha and Jaiswal, Pankaj and Chang, Jeff H and Palusa, Saiprasad G and Reddy, A S N and Megraw, Molly and Mockler, Todd C} } @article {319, title = {Paired-end analysis of transcription start sites in Arabidopsis reveals plant-specific promoter signatures.}, journal = {Plant Cell}, volume = {26}, year = {2014}, month = {2014 Jul}, pages = {2746-60}, abstract = {

Understanding plant gene promoter architecture has long been a challenge due to the lack of relevant large-scale data sets and analysis methods. Here, we present a publicly available, large-scale transcription start site (TSS) data set in plants using a high-resolution method for analysis of 5\&$\#$39; ends of mRNA transcripts. Our data set is produced using the paired-end analysis of transcription start sites (PEAT) protocol, providing millions of TSS locations from wild-type Columbia-0 Arabidopsis thaliana whole root samples. Using this data set, we grouped TSS reads into \"TSS tag clusters\" and categorized clusters into three spatial initiation patterns: narrow peak, broad with peak, and weak peak. We then designed a machine learning model that predicts the presence of TSS tag clusters with outstanding sensitivity and specificity for all three initiation patterns. We used this model to analyze the transcription factor binding site content of promoters exhibiting these initiation patterns. In contrast to the canonical notions of TATA-containing and more broad \"TATA-less\" promoters, the model shows that, in plants, the vast majority of transcription start sites are TATA free and are defined by a large compendium of known DNA sequence binding elements. We present results on the usage of these elements and provide our Plant PEAT Peaks (3PEAT) model that predicts the presence of TSSs directly from sequence.

[Link to Additional Data and Supplementary Materials]

}, keywords = {Arabidopsis, Arabidopsis Proteins, Binding Sites, Cluster Analysis, DNA, Plant, Gene Expression Regulation, Plant, Genome, Plant, Models, Genetic, Nucleotide Motifs, Plant Roots, Promoter Regions, Genetic, RNA, Messenger, RNA, Plant, Sequence Analysis, DNA, Species Specificity, TATA Box, Transcription Factors, Transcription Initiation Site}, issn = {1532-298X}, doi = {10.1105/tpc.114.125617}, author = {Morton, Taj and Petricka, Jalean and Corcoran, David L and Li, Song and Winter, Cara M and Carda, Alexa and Benfey, Philip N and Ohler, Uwe and Megraw, Molly} } @article {321, title = {The protein expression landscape of the Arabidopsis root.}, journal = {Proc Natl Acad Sci U S A}, volume = {109}, year = {2012}, month = {2012 May 1}, pages = {6811-8}, abstract = {

Because proteins are the major functional components of cells, knowledge of their cellular localization is crucial to gaining an understanding of the biology of multicellular organisms. We have generated a protein expression map of the Arabidopsis root providing the identity and cell type-specific localization of nearly 2,000 proteins. Grouping proteins into functional categories revealed unique cellular functions and identified cell type-specific biomarkers. Cellular colocalization provided support for numerous protein-protein interactions. With a binary comparison, we found that RNA and protein expression profiles are weakly correlated. We then performed peak integration at cell type-specific resolution and found an improved correlation with transcriptome data using continuous values. We performed GeLC-MS/MS (in-gel tryptic digestion followed by liquid chromatography-tandem mass spectrometry) proteomic experiments on mutants with ectopic and no root hairs, providing complementary proteomic data. Finally, among our root hair-specific proteins we identified two unique regulators of root hair development.

}, keywords = {Arabidopsis, Arabidopsis Proteins, Base Sequence, Chromatography, Liquid, DNA Primers, Gene Expression Profiling, Plant Roots, Plants, Genetically Modified, Protein Array Analysis, Protein Interaction Mapping, Proteome, Proteomics, RNA, Plant, Tandem Mass Spectrometry}, issn = {1091-6490}, doi = {10.1073/pnas.1202546109}, author = {Petricka, Jalean J and Schauer, Monica A and Megraw, Molly and Breakfield, Natalie W and Thompson, J Will and Georgiev, Stoyan and Soderblom, Erik J and Ohler, Uwe and Moseley, Martin Arthur and Grossniklaus, Ueli and Benfey, Philip N} } @article {324, title = {miRGen 2.0: a database of microRNA genomic information and regulation.}, journal = {Nucleic Acids Res}, volume = {38}, year = {2010}, month = {2010 Jan}, pages = {D137-41}, abstract = {

MicroRNAs are small, non-protein coding RNA molecules known to regulate the expression of genes by binding to the 3\&$\#$39;UTR region of mRNAs. MicroRNAs are produced from longer transcripts which can code for more than one mature miRNAs. miRGen 2.0 is a database that aims to provide comprehensive information about the position of human and mouse microRNA coding transcripts and their regulation by transcription factors, including a unique compilation of both predicted and experimentally supported data. Expression profiles of microRNAs in several tissues and cell lines, single nucleotide polymorphism locations, microRNA target prediction on protein coding genes and mapping of miRNA targets of co-regulated miRNAs on biological pathways are also integrated into the database and user interface. The miRGen database will be continuously maintained and freely available at http://www.microrna.gr/mirgen/.

}, keywords = {3{\textquoteright} Untranslated Regions, Algorithms, Animals, Cell Line, Tumor, Computational Biology, Databases, Genetic, Databases, Nucleic Acid, Humans, Information Storage and Retrieval, Internet, Mice, MicroRNAs, Polymorphism, Single Nucleotide, Software, Transcription Factors}, issn = {1362-4962}, doi = {10.1093/nar/gkp888}, author = {Alexiou, Panagiotis and Vergoulis, Thanasis and Gleditzsch, Martin and Prekas, George and Dalamagas, Theodore and Megraw, Molly and Grosse, Ivo and Sellis, Timos and Hatzigeorgiou, Artemis G} } @article {326, title = {A transcription factor affinity-based code for mammalian transcription initiation.}, journal = {Genome Res}, volume = {19}, year = {2009}, month = {2009 Apr}, pages = {644-56}, abstract = {

The recent arrival of large-scale cap analysis of gene expression (CAGE) data sets in mammals provides a wealth of quantitative information on coding and noncoding RNA polymerase II transcription start sites (TSS). Genome-wide CAGE studies reveal that a large fraction of TSS exhibit peaks where the vast majority of associated tags map to a particular location ( approximately 45\%), whereas other active regions contain a broader distribution of initiation events. The presence of a strong single peak suggests that transcription at these locations may be mediated by position-specific sequence features. We therefore propose a new model for single-peaked TSS based solely on known transcription factors (TFs) and their respective regions of positional enrichment. This probabilistic model leads to near-perfect classification results in cross-validation (auROC = 0.98), and performance in genomic scans demonstrates that TSS prediction with both high accuracy and spatial resolution is achievable for a specific but large subgroup of mammalian promoters. The interpretable model structure suggests a DNA code in which canonical sequence features such as TATA-box, Initiator, and GC content do play a significant role, but many additional TFs show distinct spatial biases with respect to TSS location and are important contributors to the accurate prediction of single-peak transcription initiation sites. The model structure also reveals that CAGE tag clusters distal from annotated gene starts have distinct characteristics compared to those close to gene 5\&$\#$39;-ends. Using this high-resolution single-peak model, we predict TSS for approximately 70\% of mammalian microRNAs based on currently available data.

[Links to Tools and Supplementary Materials]

}, keywords = {Base Composition, Databases, Genetic, DNA, Gene Expression Regulation, Genome, Human, Humans, Promoter Regions, Genetic, RNA Polymerase II, TATA Box, Transcription Factors, Transcription Initiation Site, Transcription, Genetic}, issn = {1088-9051}, doi = {10.1101/gr.085449.108}, author = {Megraw, Molly and Pereira, Fernando and Jensen, Shane T and Ohler, Uwe and Hatzigeorgiou, Artemis G} }