Selective nuclear export of specific classes of mRNA from mammalian nuclei is promoted by GANP

The nuclear phase of the gene expression pathway culminates in the export of mature messenger RNAs (mRNAs) to the cytoplasm through nuclear pore complexes. GANP (germinal- centre associated nuclear protein) promotes the transfer of mRNAs bound to the transport factor NXF1 to nuclear pore complexes. Here, we demonstrate that GANP, subunit of the TRanscription-EXport-2 (TREX-2) mRNA export complex, promotes selective nuclear export of a specific subset of mRNAs whose transport depends on NXF1. Genome-wide gene expression profiling showed that half of the transcripts whose nuclear export was impaired following NXF1 depletion also showed reduced export when GANP was depleted. GANP-dependent transcripts were highly expressed, yet short-lived, and were highly enriched in those encoding central components of the gene expression machinery such as RNA synthesis and processing factors. After injection into Xenopus oocyte nuclei, representative GANP-dependent transcripts showed faster nuclear export kinetics than representative transcripts that were not influenced by GANP depletion. We propose that GANP promotes the nuclear export of specific classes of mRNAs that may facilitate rapid changes in gene expression.

[1]  G. Hong,et al.  Nucleic Acids Research , 2015, Nucleic Acids Research.

[2]  M. Babu,et al.  Human inositol polyphosphate multikinase regulates transcript-selective nuclear mRNA export to preserve genome integrity. , 2013, Molecular cell.

[3]  F. Waharte,et al.  The human TREX-2 complex is stably associated with the nuclear pore basket , 2013, Journal of Cell Science.

[4]  M. Ono,et al.  GANP regulates recruitment of AID to immunoglobulin variable regions by modulating transcription and nucleosome occupancy , 2013, Nature Communications.

[5]  K. Neugebauer,et al.  How cells get the message: dynamic assembly and function of mRNA–protein complexes , 2013, Nature Reviews Genetics.

[6]  E. Hurt,et al.  Functional and structural characterization of the mammalian TREX-2 complex that links transcription with nuclear messenger RNA export , 2012, Nucleic acids research.

[7]  Stuart A. Wilson,et al.  TREX exposes the RNA binding domain of Nxf1 to enable mRNA export , 2012, Nature Communications.

[8]  F. Stutz,et al.  Keeping mRNPs in check during assembly and nuclear export , 2011, Nature Reviews Molecular Cell Biology.

[9]  Ed Hurt,et al.  Linking gene regulation to mRNA production and export. , 2011, Current opinion in cell biology.

[10]  T. Tamura,et al.  Identification of mRNAs that are spliced but not exported to the cytoplasm in the absence of THOC5 in mouse embryo fibroblasts. , 2011, RNA.

[11]  M. Selbach,et al.  Global quantification of mammalian gene expression control , 2011, Nature.

[12]  M. Mann,et al.  Defining the transcriptome and proteome in three functionally different human cell lines , 2010, Molecular systems biology.

[13]  M. Stewart Nuclear export of mRNA. , 2010, Trends in biochemical sciences.

[14]  Y. Shav-Tal,et al.  Dynamics of single mRNP nucleocytoplasmic transport and export through the nuclear pore in living cells , 2010, Nature Cell Biology.

[15]  E. Kondo,et al.  Germinal center‐associated nuclear protein (GANP) is involved in mRNA export of Shugoshin‐1 required for centromere cohesion and in sister‐chromatid exchange , 2010, Genes to cells : devoted to molecular & cellular mechanisms.

[16]  J. Thyberg,et al.  Protein Tpr is required for establishing nuclear pore-associated zones of heterochromatin exclusion , 2010, The EMBO journal.

[17]  V. Wickramasinghe,et al.  mRNA Export from Mammalian Cell Nuclei Is Dependent on GANP , 2010, Current Biology.

[18]  Sarath Chandra Janga,et al.  Dissecting the expression dynamics of RNA-binding proteins in posttranscriptional regulatory networks , 2009, Proceedings of the National Academy of Sciences.

[19]  Caroline C. Friedel,et al.  Conserved principles of mammalian transcriptional regulation revealed by RNA half-life , 2009, Nucleic acids research.

[20]  J. Bähler,et al.  Rapidly regulated genes are intron poor. , 2008, Trends in genetics : TIG.

[21]  Ed Hurt,et al.  Exporting RNA from the nucleus to the cytoplasm , 2007, Nature Reviews Molecular Cell Biology.

[22]  Elizabeth J. Tran,et al.  Dynamic Nuclear Pore Complexes: Life on the Edge , 2006, Cell.

[23]  Jean-Christophe Olivo-Marin,et al.  SAGA interacting factors confine sub-diffusion of transcribed genes to the nuclear envelope , 2006, Nature.

[24]  F. Hediger,et al.  Nuclear pore association confers optimal expression levels for an inducible yeast gene , 2006, Nature.

[25]  Seiji Masuda,et al.  Recruitment of the human TREX complex to mRNA during splicing. , 2005, Genes & development.

[26]  H. Hieronymus,et al.  A systems view of mRNP biology. , 2004, Genes & development.

[27]  T. Köcher,et al.  Genome-wide analysis of mRNAs regulated by the THO complex in Drosophila melanogaster , 2004, Nature Structural &Molecular Biology.

[28]  Gordon K Smyth,et al.  Linear Models and Empirical Bayes Methods for Assessing Differential Expression in Microarray Experiments , 2004, Statistical applications in genetics and molecular biology.

[29]  Oreto Antúnez,et al.  Sus1, a Functional Component of the SAGA Histone Acetylase Complex and the Nuclear Pore-Associated mRNA Export Machinery , 2004, Cell.

[30]  Brad T. Sherman,et al.  DAVID: Database for Annotation, Visualization, and Integrated Discovery , 2003, Genome Biology.

[31]  E. Izaurralde,et al.  Genome‐wide analysis of nuclear mRNA export pathways in Drosophila , 2003, The EMBO journal.

[32]  J. Steitz,et al.  SR splicing factors serve as adapter proteins for TAP-dependent mRNA export. , 2003, Molecular cell.

[33]  H. Hieronymus,et al.  Genome-wide analysis of RNA–protein interactions illustrates specificity of the mRNA export machinery , 2003, Nature Genetics.

[34]  Tamás Fischer,et al.  The mRNA export machinery requires the novel Sac3p–Thp1p complex to dock at the nucleoplasmic entrance of the nuclear pores , 2002, The EMBO journal.

[35]  D. Weil,et al.  In Vivo Kinetics of mRNA Splicing and Transport in Mammalian Cells , 2002, Molecular and Cellular Biology.

[36]  Cristian I. Castillo-Davis,et al.  Selection for short introns in highly expressed genes , 2002, Nature Genetics.

[37]  M. Rode,et al.  Nuclear Export of mRNA by TAP/NXF1 Requires Two Nucleoporin-Binding Sites but Not p15 , 2002, Molecular and Cellular Biology.

[38]  S. Tenenbaum,et al.  Eukaryotic mRNPs may represent posttranscriptional operons. , 2002, Molecular cell.

[39]  S. Shibata,et al.  Nucleocytoplasmic transport of proteins and poly(A)+ RNA in reconstituted Tpr‐less nuclei in living mammalian cells , 2002, Genes to cells : devoted to molecular & cellular mechanisms.

[40]  E. Hurt,et al.  Structure of the C-terminal FG-nucleoporin binding domain of Tap/NXF1 , 2002, Nature Structural Biology.

[41]  Ed Hurt,et al.  A Conserved mRNA Export Machinery Coupled to pre-mRNA Splicing , 2002, Cell.

[42]  S. Dudoit,et al.  Normalization for cDNA microarray data: a robust composite method addressing single and multiple slide systematic variation. , 2002, Nucleic acids research.

[43]  G. Blobel,et al.  The nucleoporin Nup98 associates with the intranuclear filamentous protein network of TPR , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[44]  M. Suyama,et al.  TAP (NXF1) Belongs to a Multigene Family of Putative RNA Export Factors with a Conserved Modular Architecture , 2000, Molecular and Cellular Biology.

[45]  E. Hurt,et al.  The protein Aly links pre-messenger-RNA splicing to nuclear export in metazoans , 2000, Nature.

[46]  A. Podtelejnikov,et al.  The Mex67p‐mediated nuclear mRNA export pathway is conserved from yeast to human , 1999, The EMBO journal.

[47]  R. Singer,et al.  Movement of nuclear poly(A) RNA throughout the interchromatin space in living cells , 1999, Current Biology.

[48]  Michael P. Rout,et al.  Proteins Connecting the Nuclear Pore Complex with the Nuclear Interior , 1999, The Journal of cell biology.

[49]  S. Doxsey,et al.  Functional Analysis of Tpr: Identification of Nuclear Pore Complex Association and Nuclear Localization Domains and a Role in mRNA Export , 1998, The Journal of cell biology.

[50]  V. Cordes,et al.  Identification of Protein p270/Tpr as a Constitutive Component of the Nuclear Pore Complex–attached Intranuclear Filaments , 1997, The Journal of cell biology.

[51]  I. Mattaj,et al.  Monomethylated cap structures facilitate RNA export from the nucleus , 1990, Cell.

[52]  Brad T. Sherman,et al.  Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources , 2008, Nature Protocols.

[53]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .