ASAP, a Novel Protein Complex Involved in RNA Processing and Apoptosis

ABSTRACT Different isoforms of a protein complex termed the apoptosis- and splicing-associated protein (ASAP) were isolated from HeLa cell extract. ASAP complexes are composed of the polypeptides SAP18 and RNPS1 and different isoforms of the Acinus protein. While Acinus had previously been implicated in apoptosis and was recently identified as a component of the spliceosome, RNPS1 has been described as a general activator of RNA processing. Addition of ASAP isoforms to in vitro splicing reactions inhibits RNA processing mediated by ASF/SF2, by SC35, or by RNPS1. Additionally, microinjection of ASAP complexes into mammalian cells resulted in acceleration of cell death. Importantly, after induction of apoptosis the ASAP complex disassembles. Taken together, our results suggest an important role for the ASAP complexes in linking RNA processing and apoptosis.

[1]  Steven P. Gygi,et al.  Comprehensive proteomic analysis of the human spliceosome , 2002, Nature.

[2]  J. Gottesfeld,et al.  Cyclin L Is an RS Domain Protein Involved in Pre-mRNA Splicing* , 2002, The Journal of Biological Chemistry.

[3]  Bosiljka Tasic,et al.  Alternative pre-mRNA splicing and proteome expansion in metazoans , 2002, Nature.

[4]  J. Bishop,et al.  Suppressor of Fused represses Gli-mediated transcription by recruiting the SAP18-mSin3 corepressor complex , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Y. Hannun,et al.  De Novo Ceramide Regulates the Alternative Splicing of Caspase 9 and Bcl-x in A549 Lung Adenocarcinoma Cells , 2002, The Journal of Biological Chemistry.

[6]  T. Maniatis,et al.  An extensive network of coupling among gene expression machines , 2002, Nature.

[7]  G. Orphanides,et al.  A Unified Theory of Gene Expression , 2002, Cell.

[8]  S. Hyman,et al.  Dopamine and Glutamate Induce Distinct Striatal Splice Forms of Ania-6, an RNA Polymerase II-Associated Cyclin , 2001, Neuron.

[9]  J. Steitz,et al.  Communication of the Position of Exon-Exon Junctions to the mRNA Surveillance Machinery by the Protein RNPS1 , 2001, Science.

[10]  H. Le Hir,et al.  The exon–exon junction complex provides a binding platform for factors involved in mRNA export and nonsense‐mediated mRNA decay , 2001, The EMBO journal.

[11]  H. Le Hir,et al.  The spliceosome deposits multiple proteins 20–24 nucleotides upstream of mRNA exon–exon junctions , 2000, The EMBO journal.

[12]  B. Graveley.  Sorting out the complexity of SR protein functions. , 2000, RNA.

[13]  J. Manley,et al.  RNA polymerase II and the integration of nuclear events. , 2000, Genes & development.

[14]  T Misteli,et al.  Cell biology of transcription and pre-mRNA splicing: nuclear architecture meets nuclear function. , 2000, Journal of cell science.

[15]  G. Häcker.  The morphology of apoptosis , 2000, Cell and Tissue Research.

[16]  E. Koonin,et al.  SAP - a putative DNA-binding motif involved in chromosomal organization. , 2000, Trends in biochemical sciences.

[17]  P. Tempst,et al.  Tuning of an electrospray ionization source for maximum peptide-ion transmission into a mass spectrometer. , 2000, Analytical chemistry.

[18]  T. Pawson,et al.  The Protein Kinase Clk/Sty Directly Modulates SR Protein Activity: Both Hyper- and Hypophosphorylation Inhibit Splicing , 1999, Molecular and Cellular Biology.

[19]  N. Imamoto,et al.  Acinus is a caspase-3-activated protein required for apoptotic chromatin condensation , 1999, Nature.

[20]  Michael Q. Zhang,et al.  Purification and characterization of human RNPS1: a general activator of pre‐mRNA splicing , 1999, The EMBO journal.

[21]  Jane Y. Wu,et al.  Alternative splicing and programmed cell death. , 1999, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[22]  S. Carr,et al.  Examination of micro-tip reversed-phase liquid chromatographic extraction of peptide pools for mass spectrometric analysis. , 1998, Journal of chromatography. A.

[23]  D. Reinberg,et al.  The Dermatomyositis-Specific Autoantigen Mi2 Is a Component of a Complex Containing Histone Deacetylase and Nucleosome Remodeling Activities , 1998, Cell.

[24]  D. Reinberg,et al.  SAP30, a novel protein conserved between human and yeast, is a component of a histone deacetylase complex. , 1998, Molecular cell.

[25]  P. Loyer,et al.  The RNP protein, RNPS1, associates with specific isoforms of the p34cdc2-related PITSLRE protein kinase in vivo. , 1998, Journal of cell science.

[26]  D. Reinberg,et al.  Histone Deacetylases and SAP18, a Novel Polypeptide, Are Components of a Human Sin3 Complex , 1997, Cell.

[27]  J. Manley,et al.  SR proteins and splicing control. , 1996, Genes & development.

[28]  D. Higgs,et al.  Mutations in a putative global transcriptional regulator cause X-linked mental retardation with α-thalassemia (ATR-X syndrome) , 1995, Cell.

[29]  P. Højrup,et al.  Use of mass spectrometric molecular weight information to identify proteins in sequence databases. , 1993, Biological mass spectrometry.

[30]  A. Strasser,et al.  Apoptosis signaling. , 2000, Annual review of biochemistry.

[31]  S H Kaufmann,et al.  Mammalian caspases: structure, activation, substrates, and functions during apoptosis. , 1999, Annual review of biochemistry.

[32]  A. J. Lopez,et al.  Alternative splicing of pre-mRNA: developmental consequences and mechanisms of regulation. , 1998, Annual review of genetics.

[33]  V. Kidd,et al.  Proteolytic activities that mediate apoptosis. , 1998, Annual review of physiology.

[34]  A. Krämer,et al.  The structure and function of proteins involved in mammalian pre-mRNA splicing. , 1996, Annual review of biochemistry.

[35]  D. Spector,et al.  Macromolecular domains within the cell nucleus. , 1993, Annual review of cell biology.