title: The Composition of the Arabidopsis RNA Polymerase II Transcript Elongation Complex Reveals the Interplay between Elongation and mRNA Processing Factors.
abstract: Transcript elongation factors (TEFs) are a heterogeneous group of proteins that control the efficiency of transcript elongation of subsets of genes by RNA polymerase II (RNAPII) in the chromatin context. Using reciprocal tagging in combination with affinity purification and mass spectrometry, we demonstrate that inArabidopsis thaliana, the TEFs SPT4/SPT5, SPT6, FACT, PAF1-C, and TFIIS copurified with each other and with elongating RNAPII, while P-TEFb was not among the interactors. Additionally, NAP1 histone chaperones, ATP-dependent chromatin remodeling factors, and some histone-modifying enzymes including Elongator were repeatedly found associated with TEFs. Analysis of double mutant plants defective in different combinations of TEFs revealed genetic interactions between genes encoding subunits of PAF1-C, FACT, and TFIIS, resulting in synergistic/epistatic effects on plant growth/development. Analysis of subnuclear localization, gene expression, and chromatin association did not provide evidence for an involvement of the TEFs in transcription by RNAPI (or RNAPIII). Proteomics analyses also revealed multiple interactions between the transcript elongation complex and factors involved in mRNA splicing and polyadenylation, including an association of PAF1-C with the polyadenylation factor CstF. Therefore, the RNAPII transcript elongation complex represents a platform for interactions among different TEFs, as well as for coordinating ongoing transcription with mRNA processing.
authors.affiliation: Department of Cell Biology and Plant Biochemistry, Biochemistry Center, University of Regensburg, D-93053 Regensburg, Germany.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Antosz
authors.affiliation: Department of Cell Biology and Plant Biochemistry, Biochemistry Center, University of Regensburg, D-93053 Regensburg, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Pfab
authors.affiliation: Department of Cell Biology and Plant Biochemistry, Biochemistry Center, University of Regensburg, D-93053 Regensburg, Germany.
authors.forename: Hans F
authors.identifier: http://orcid.org/0000-0002-0007-0485
authors.initials: HF
authors.lastname: Ehrnsberger
authors.affiliation: Department of Cell Biology and Plant Biochemistry, Biochemistry Center, University of Regensburg, D-93053 Regensburg, Germany.
authors.forename: Philipp
authors.identifier: http://orcid.org/0000-0003-3286-0905
authors.initials: P
authors.lastname: Holzinger
authors.affiliation: Department of Cell Biology and Plant Biochemistry, Biochemistry Center, University of Regensburg, D-93053 Regensburg, Germany.
authors.forename: Karin
authors.initials: K
authors.lastname: Köllen
authors.affiliation: Department of Cell Biology and Plant Biochemistry, Biochemistry Center, University of Regensburg, D-93053 Regensburg, Germany.
authors.forename: Simon A
authors.identifier: http://orcid.org/0000-0001-9013-1802
authors.initials: SA
authors.lastname: Mortensen
authors.affiliation: Department for Biochemistry I, Biochemistry Center, University of Regensburg, D-93053 Regensburg, Germany.
authors.forename: Astrid
authors.initials: A
authors.lastname: Bruckmann
authors.affiliation: Department for Biochemistry III, Biochemistry Center, University of Regensburg, D-93053 Regensburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Schubert
authors.affiliation: Department for Biochemistry III, Biochemistry Center, University of Regensburg, D-93053 Regensburg, Germany.
authors.forename: Gernot
authors.identifier: http://orcid.org/0000-0002-8232-1179
authors.initials: G
authors.lastname: Längst
authors.affiliation: Department for Biochemistry III, Biochemistry Center, University of Regensburg, D-93053 Regensburg, Germany.
authors.forename: Joachim
authors.identifier: http://orcid.org/0000-0002-7817-6095
authors.initials: J
authors.lastname: Griesenbeck
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben, D-06466 Stadt Seeland, Germany.
authors.forename: Veit
authors.identifier: http://orcid.org/0000-0002-3072-0485
authors.initials: V
authors.lastname: Schubert
authors.affiliation: Department of Cell Biology and Plant Biochemistry, Biochemistry Center, University of Regensburg, D-93053 Regensburg, Germany marion.grasser@ur.de klaus.grasser@ur.de.
authors.forename: Marion
authors.initials: M
authors.lastname: Grasser
authors.affiliation: Department of Cell Biology and Plant Biochemistry, Biochemistry Center, University of Regensburg, D-93053 Regensburg, Germany marion.grasser@ur.de klaus.grasser@ur.de.
authors.forename: Klaus D
authors.identifier: http://orcid.org/0000-0002-7080-5520
authors.initials: KD
authors.lastname: Grasser
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D056488:Histone Chaperones / Q000235:genetics / Q000378:metabolism; D040901:Proteomics; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000235:genetics; D035602:Transcriptional Elongation Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D056488:Histone Chaperones; D012333:RNA, Messenger; D018749:RNA, Plant; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; C018648:transcription factor S-II; D012319:RNA Polymerase II; D000251:Adenosine Triphosphatases; C498859:NAP1 protein, Arabidopsis