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      Promoter‐pervasive transcription causes RNA polymerase II pausing to boost DOG1 expression in response to salt

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          Abstract

          Eukaryotic genomes are pervasively transcribed by RNA polymerase II. Yet, the molecular and biological implications of such a phenomenon are still largely puzzling. Here, we describe noncoding RNA transcription upstream of the Arabidopsis thaliana DOG1 gene, which governs salt stress responses and is a key regulator of seed dormancy. We find that expression of the DOG1 gene is induced by salt stress, thereby causing a delay in seed germination. We uncover extensive transcriptional activity on the promoter of the DOG1 gene, which produces a variety of lncRNAs. These lncRNAs, named PUPPIES, are co‐directionally transcribed and extend into the DOG1 coding region. We show that PUPPIES RNAs respond to salt stress and boost DOG1 expression, resulting in delayed germination. This positive role of pervasive PUPPIES transcription on DOG1 gene expression is associated with augmented pausing of RNA polymerase II, slower transcription and higher transcriptional burst size. These findings highlight the positive role of upstream co‐directional transcription in controlling transcriptional dynamics of downstream genes.

          Abstract

          PUPPIES lncRNAs transcription from the promoter of seed dormancy regulator DOG1 augments pausing of RNA polymerase II and delays germination.

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          Most cited references106

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          STAR: ultrafast universal RNA-seq aligner.

          Accurate alignment of high-throughput RNA-seq data is a challenging and yet unsolved problem because of the non-contiguous transcript structure, relatively short read lengths and constantly increasing throughput of the sequencing technologies. Currently available RNA-seq aligners suffer from high mapping error rates, low mapping speed, read length limitation and mapping biases. To align our large (>80 billon reads) ENCODE Transcriptome RNA-seq dataset, we developed the Spliced Transcripts Alignment to a Reference (STAR) software based on a previously undescribed RNA-seq alignment algorithm that uses sequential maximum mappable seed search in uncompressed suffix arrays followed by seed clustering and stitching procedure. STAR outperforms other aligners by a factor of >50 in mapping speed, aligning to the human genome 550 million 2 × 76 bp paired-end reads per hour on a modest 12-core server, while at the same time improving alignment sensitivity and precision. In addition to unbiased de novo detection of canonical junctions, STAR can discover non-canonical splices and chimeric (fusion) transcripts, and is also capable of mapping full-length RNA sequences. Using Roche 454 sequencing of reverse transcription polymerase chain reaction amplicons, we experimentally validated 1960 novel intergenic splice junctions with an 80-90% success rate, corroborating the high precision of the STAR mapping strategy. STAR is implemented as a standalone C++ code. STAR is free open source software distributed under GPLv3 license and can be downloaded from http://code.google.com/p/rna-star/.
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            Cutadapt removes adapter sequences from high-throughput sequencing reads

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              BEDTools: a flexible suite of utilities for comparing genomic features

              Motivation: Testing for correlations between different sets of genomic features is a fundamental task in genomics research. However, searching for overlaps between features with existing web-based methods is complicated by the massive datasets that are routinely produced with current sequencing technologies. Fast and flexible tools are therefore required to ask complex questions of these data in an efficient manner. Results: This article introduces a new software suite for the comparison, manipulation and annotation of genomic features in Browser Extensible Data (BED) and General Feature Format (GFF) format. BEDTools also supports the comparison of sequence alignments in BAM format to both BED and GFF features. The tools are extremely efficient and allow the user to compare large datasets (e.g. next-generation sequencing data) with both public and custom genome annotation tracks. BEDTools can be combined with one another as well as with standard UNIX commands, thus facilitating routine genomics tasks as well as pipelines that can quickly answer intricate questions of large genomic datasets. Availability and implementation: BEDTools was written in C++. Source code and a comprehensive user manual are freely available at http://code.google.com/p/bedtools Contact: aaronquinlan@gmail.com; imh4y@virginia.edu Supplementary information: Supplementary data are available at Bioinformatics online.
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                Author and article information

                Contributors
                miguel-coelho@hotmail.com
                sswiez@ibb.waw.pl
                Journal
                EMBO J
                EMBO J
                10.1002/(ISSN)1460-2075
                EMBJ
                embojnl
                The EMBO Journal
                John Wiley and Sons Inc. (Hoboken )
                0261-4189
                1460-2075
                27 January 2023
                March 2023
                27 January 2023
                : 42
                : 5 ( doiID: 10.1002/embj.v42.5 )
                : e112443
                Affiliations
                [ 1 ] Laboratory of Seeds Molecular Biology, Institute of Biochemistry and Biophysics Polish Academy of Sciences Warsaw Poland
                [ 2 ] Laboratory of Functional and Structural Genomics, Centre of New Technologies University of Warsaw Warsaw Poland
                [ 3 ] Laboratory of Bioinformatics and Computational Genomics, Faculty of Mathematics and Information Science Warsaw University of Technology Warsaw Poland
                Author notes
                [*] [* ] Corresponding author. Tel: +48 22 592 57 25; E‐mail: miguel-coelho@ 123456hotmail.com

                Corresponding author. Tel: +48 22 592 57 25; E‐mail: sswiez@ 123456ibb.waw.pl

                Author information
                https://orcid.org/0000-0002-1383-0885
                https://orcid.org/0000-0002-5714-1993
                https://orcid.org/0000-0002-9795-5588
                https://orcid.org/0000-0001-8220-9649
                https://orcid.org/0000-0001-9619-9664
                https://orcid.org/0000-0001-5382-4079
                https://orcid.org/0000-0003-4480-314X
                https://orcid.org/0000-0002-3840-7610
                https://orcid.org/0000-0002-2007-2779
                Article
                EMBJ2022112443
                10.15252/embj.2022112443
                9975946
                36705062
                2f85c50a-b60b-4fca-8932-7bfe92141d73
                © 2023 The Authors. Published under the terms of the CC BY 4.0 license

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 02 January 2023
                : 23 August 2022
                : 09 January 2023
                Page count
                Figures: 13, Tables: 1, Pages: 27, Words: 21018
                Funding
                Funded by: COST , doi 10.13039/501100000921;
                Award ID: CA16212
                Funded by: Fundacja na rzecz Nauki Polskiej (FNP) , doi 10.13039/501100001870;
                Award ID: TEAM POIR.04.04.00‐00‐3C97/16
                Funded by: Narodowe Centrum Nauki (NCN) , doi 10.13039/501100004281;
                Award ID: SONATA BIS UMO‐2018/30/E/NZ1/00354
                Award ID: OPUS UMO‐2018/31/B/NZ3/03363
                Award ID: NCN 2019/35/O/ST6/02484
                Award ID: 2020/37/B/NZ2/03757
                Award ID: UMO‐2021/41/B/NZ3/02605
                Funded by: Fundação para a Ciência e Tecnologia (FCT) , doi 10.13039/501100005856;
                Award ID: SFRH/BD/145264/2019
                Funded by: Warsaw University of Technology , doi 10.13039/501100004421;
                Categories
                Article
                Articles
                Custom metadata
                2.0
                01 March 2023
                Converter:WILEY_ML3GV2_TO_JATSPMC version:6.2.5 mode:remove_FC converted:01.03.2023

                Molecular biology
                long noncoding rnas,rna polymerase ii pausing,salt stress,transcriptional dynamics,chromatin, transcription & genomics,plant biology

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