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      Mud2 functions in transcription by recruiting the Prp19 and TREX complexes to transcribed genes

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          Abstract

          The different steps of gene expression are intimately linked to coordinate and regulate this complex process. During transcription, numerous RNA-binding proteins are already loaded onto the nascent mRNA and package the mRNA into a messenger ribonucleoprotein particle (mRNP). These RNA-binding proteins are often also involved in other steps of gene expression than mRNA packaging. For example, TREX functions in transcription, mRNP packaging and nuclear mRNA export. Previously, we showed that the Prp19 splicing complex (Prp19C) is needed for efficient transcription as well as TREX occupancy at transcribed genes. Here, we show that the splicing factor Mud2 interacts with Prp19C and is needed for Prp19C occupancy at transcribed genes in Saccharomyces cerevisiae. Interestingly, Mud2 is not only recruited to intron-containing but also to intronless genes indicating a role in transcription. Indeed, we show for the first time that Mud2 functions in transcription. Furthermore, these functions of Mud2 are likely evolutionarily conserved as Mud2 is also recruited to an intronless gene and interacts with Prp19C in Drosophila melanogaster. Taken together, we classify Mud2 as a novel transcription factor that is necessary for the recruitment of mRNA-binding proteins to the transcription machinery. Thus, Mud2 is a multifunctional protein important for transcription, splicing and most likely also mRNP packaging.

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          Most cited references65

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          The tandem affinity purification (TAP) method: a general procedure of protein complex purification.

          Identification of components present in biological complexes requires their purification to near homogeneity. Methods of purification vary from protein to protein, making it impossible to design a general purification strategy valid for all cases. We have developed the tandem affinity purification (TAP) method as a tool that allows rapid purification under native conditions of complexes, even when expressed at their natural level. Prior knowledge of complex composition or function is not required. The TAP method requires fusion of the TAP tag, either N- or C-terminally, to the target protein of interest. Starting from a relatively small number of cells, active macromolecular complexes can be isolated and used for multiple applications. Variations of the method to specifically purify complexes containing two given components or to subtract undesired complexes can easily be implemented. The TAP method was initially developed in yeast but can be successfully adapted to various organisms. Its simplicity, high yield, and wide applicability make the TAP method a very useful procedure for protein purification and proteome exploration. Copyright 2001 Academic Press.
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            The code and beyond: transcription regulation by the RNA polymerase II carboxy-terminal domain

            The carboxy-terminal domain (CTD) of RNA polymerase II is a repetitive and unstructured domain that is dynamically modified by post-translational modifications, which collectively constitute the 'CTD code'. Recent studies have revealed how CTD function is also promoted by phase separation in the presence of other low-complexity domains.
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              How cells get the message: dynamic assembly and function of mRNA-protein complexes.

              mRNA is packaged into ribonucleoprotein particles called mRNPs. A multitude of RNA-binding proteins as well as a host of associated proteins participate in the fate of mRNA from transcription and processing in the nucleus to translation and decay in the cytoplasm. Methodological innovations in cell biology and genome-wide high-throughput approaches have revealed an unexpected diversity of mRNA-associated proteins and unforeseen interconnections between mRNA-processing steps. Recent insights into mRNP formation in vivo have also highlighted the importance of mRNP packaging, which can sort RNAs on the basis of their length and determine mRNA fate through alternative mRNP assembly, processing and export pathways.
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                Author and article information

                Journal
                Nucleic Acids Res
                Nucleic Acids Res
                nar
                Nucleic Acids Research
                Oxford University Press
                0305-1048
                1362-4962
                12 October 2018
                24 July 2018
                24 July 2018
                : 46
                : 18
                : 9749-9763
                Affiliations
                [1 ]Institute of Biochemistry, Justus Liebig University, Giessen 35392, Germany
                [2 ]Institute of Gene Biology, Russian Academy of Sciences, Moscow 119334, Russia
                [3 ]Institute of Molecular Medicine, Martin-Luther-University Halle Wittenberg, Halle 06120, Germany
                [4 ]Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, Russia
                Author notes
                To whom correspondence should be addressed. Tel: +49 641 99 35400; Fax: +49 641 99 35409; Email: katja.straesser@ 123456chemie.bio.uni-giessen.de
                Article
                gky640
                10.1093/nar/gky640
                6182176
                30053068
                71837574-88d5-4b52-a5b3-12df41c631b7
                © The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@ 123456oup.com

                History
                : 20 July 2018
                : 02 July 2018
                : 07 September 2017
                Page count
                Pages: 15
                Funding
                Funded by: German Research Foundation 10.13039/501100001659
                Award ID: STR697/2-1
                Award ID: STR697/3-1
                Award ID: GRK1591
                Funded by: Russian Foundation for Basic Research 10.13039/501100002261
                Award ID: 16-34-01317
                Funded by: program of fundamental research for state academies
                Award ID: 01201363822
                Categories
                RNA and RNA-protein complexes

                Genetics
                Genetics

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