Integration of Proteomics and Genomics in Platelets

Platelets, while anucleate, contain RNA, some of which is translated into protein upon activation. Hypothesising that the platelet proteome is reflected in the transcriptome, we identified 82 proteins secreted from activated platelets and compared these, as well as published proteomic data, to the transcriptional profile. We also compared the transcriptome of platelets to other tissues to identify platelet-specific genes and used ontology to determine gene categories over-represented in platelets. RNA was isolated from highly pure platelet preparations for hybridization to Affymetrix oligonucleotide arrays. We identified 2,928 distinct messages as being present in platelets. The platelet transcriptome was compared with the proteome by relating both to UniGene clusters. Platelet proteomic data correlated well with the transcriptome, with 69% of secreted proteins detectable at the mRNA level, and similar concordance was obtained using two published datasets. While many of the most abundant mRNAs are for known platelet proteins, messages were detected for proteins not previously reported in platelets. Some of these may represent residual megakaryocyte messages; however, proteomic analysis confirmed the expression of many previously unreported genes in platelets. Transcripts for well-described platelet proteins are among the most platelet-specific messages. Ontological categories related to signal transduction, receptors, ion channels, and membranes are over-represented in platelets, while categories involved in protein synthesis are depleted. Despite the absence of gene transcription, the platelet proteome is mirrored in the transcriptome. Conversely, transcriptional analysis predicts the presence of novel proteins in the platelet. Transcriptional analysis is relevant to platelet biology, providing insights into platelet function and the mechanisms of platelet disorders.

[1]  G. Cagney,et al.  Characterization of the proteins released from activated platelets leads to localization of novel platelet proteins in human atherosclerotic lesions. , 2004, Blood.

[2]  M. Gerstein,et al.  Comparing protein abundance and mRNA expression levels on a genomic scale , 2003, Genome Biology.

[3]  E. Topol,et al.  Proteomic approach to coronary atherosclerosis shows ferritin light chain as a significant marker: evidence consistent with iron hypothesis in atherosclerosis. , 2003, Physiological genomics.

[4]  P. Perrotta,et al.  Transcript profiling of human platelets using microarray and serial analysis of gene expression. , 2003, Blood.

[5]  M. Gerstein,et al.  Genomic and proteomic analysis of the myeloid differentiation program: global analysis of gene expression during induced differentiation in the MPRO cell line. , 2002, Blood.

[6]  Michael Hecker,et al.  Transcriptome and Proteome Analysis of Bacillus subtilis Gene Expression Modulated by Amino Acid Availability , 2002, Journal of bacteriology.

[7]  Martin Vingron,et al.  Variance stabilization applied to microarray data calibration and to the quantification of differential expression , 2002, ISMB.

[8]  D. Fitzgerald,et al.  Identification of the phosphotyrosine proteome from thrombin activated platelets , 2002, Proteomics.

[9]  H. Rammensee,et al.  Human platelets express heat shock protein receptors and regulate dendritic cell maturation. , 2002, Blood.

[10]  M. Hecker,et al.  Bacillus subtilis functional genomics: global characterization of the stringent response by proteome and transcriptome analysis , 2002, Journal of bacteriology.

[11]  Mark Gerstein,et al.  Analysis of mRNA expression and protein abundance data: an approach for the comparison of the enrichment of features in the cellular population of proteins and transcripts , 2002, Bioinform..

[12]  David E. Misek,et al.  Discordant Protein and mRNA Expression in Lung Adenocarcinomas * , 2002, Molecular & Cellular Proteomics.

[13]  A. Orth,et al.  Large-scale analysis of the human and mouse transcriptomes , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Lucila Ohno-Machado,et al.  Analysis of matched mRNA measurements from two different microarray technologies , 2002, Bioinform..

[15]  R. Dwek,et al.  Towards complete analysis of the platelet proteome , 2002, Proteomics.

[16]  F. Campagne,et al.  TissueInfo: high-throughput identification of tissue expression profiles and specificity. , 2001, Nucleic acids research.

[17]  J. Yates,et al.  An automated multidimensional protein identification technology for shotgun proteomics. , 2001, Analytical chemistry.

[18]  A. Weyrich,et al.  Integrins Regulate the Intracellular Distribution of Eukaryotic Initiation Factor 4E in Platelets , 2001, The Journal of Biological Chemistry.

[19]  J. Platt,et al.  PLATELET-MEDIATED ACTIVATION OF ENDOTHELIAL CELLS: IMPLICATIONS FOR THE PATHOGENESIS OF TRANSPLANT REJECTION1 , 2001, Transplantation.

[20]  D. Dixon,et al.  Activated platelets mediate inflammatory signaling by regulated interleukin 1β synthesis , 2001, The Journal of cell biology.

[21]  J. Yates,et al.  Large-scale analysis of the yeast proteome by multidimensional protein identification technology , 2001, Nature Biotechnology.

[22]  K. Kobayashi,et al.  Combined transcriptome and proteome analysis as a powerful approach to study genes under glucose repression in Bacillus subtilis. , 2001, Nucleic acids research.

[23]  P. Carmeliet,et al.  Deficiency or inhibition of Gas6 causes platelet dysfunction and protects mice against thrombosis , 2001, Nature Medicine.

[24]  N. Copeland,et al.  A mutation in Rab27a causes the vesicle transport defects observed in ashen mice. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[25]  H. Meyer,et al.  Identification of platelet proteins separated by two‐dimensional gel electrophoresis and analyzed by matrix assisted laser desorption/ionization‐time of flight‐mass spectrometry and detection of tyrosine‐phosphorylated proteins , 2000, Electrophoresis.

[26]  R. Pawankar,et al.  Functional expression of the high affinity receptor for IgE (FcepsilonRI) in human platelets and its' intracellular expression in human megakaryocytes. , 1999, Blood.

[27]  S. Gygi,et al.  Correlation between Protein and mRNA Abundance in Yeast , 1999, Molecular and Cellular Biology.

[28]  D. Dixon,et al.  Integrin-dependent Control of Translation: Engagement of Integrin αIIbβ3 Regulates Synthesis of Proteins in Activated Human Platelets , 1999, The Journal of cell biology.

[29]  O. Bagasra,et al.  Molecular Cloning of Platelet Factor XI, an Alternative Splicing Product of the Plasma Factor XI Gene* , 1998, The Journal of Biological Chemistry.

[30]  D. Dixon,et al.  Signal-dependent translation of a regulatory protein, Bcl-3, in activated human platelets. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[31]  T. Perneger What's wrong with Bonferroni adjustments , 1998, BMJ.

[32]  L. Wodicka,et al.  Genome-wide expression monitoring in Saccharomyces cerevisiae , 1997, Nature Biotechnology.

[33]  C. Cerletti,et al.  Thrombin-activated Human Platelets Release two NAP-2 Variants that Stimulate Polymorphonuclear Leukocytes , 1996, Thrombosis and Haemostasis.

[34]  J. Cawley,et al.  Platelets prime PMN via released PF4: mechanism of priming and synergy with GM‐CSF , 1995, British journal of haematology.

[35]  C. Power,et al.  Chemokine and chemokine receptor mRNA expression in human platelets. , 1995, Cytokine.

[36]  J. Yates,et al.  An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database , 1994, Journal of the American Society for Mass Spectrometry.

[37]  A. Karniguian,et al.  Identification of small GTP-binding rab proteins in human platelets: thrombin-induced phosphorylation of rab3B, rab6, and rab8 proteins. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[38]  Y. Xu,et al.  Molecular cloning and expression of a cDNA encoding a novel isoenzyme of protein kinase C (nPKC). A new member of the nPKC family expressed in skeletal muscle, megakaryoblastic cells, and platelets. , 1993, The Journal of biological chemistry.

[39]  C. Dinarello,et al.  Activated platelets induce endothelial secretion of interleukin-8 in vitro via an interleukin-1-mediated event. , 1993, Blood.

[40]  B. Konkle,et al.  Plasminogen activator inhibitor-1 mRNA is expressed in platelets and megakaryocytes and the megakaryoblastic cell line CHRF-288. , 1993, Arteriosclerosis and thrombosis : a journal of vascular biology.

[41]  M. Feldmann,et al.  Platelet-derived interleukin 1 induces human endothelial adhesion molecule expression and cytokine production , 1991, The Journal of experimental medicine.

[42]  D. Schümperli,et al.  Regulation of histone mRNA in the unperturbed cell cycle: evidence suggesting control at two posttranscriptional steps , 1991, Molecular and cellular biology.

[43]  W. Vainchenker,et al.  Biosynthesis of major platelet proteins in human blood platelets. , 1987, European journal of biochemistry.

[44]  F. J. Morgan,et al.  In vitro synthesis of low molecular weight proteins in human platelets: absence of labelled release products. , 1984, Thrombosis research.

[45]  Joshua E. Elias,et al.  Evaluation of multidimensional chromatography coupled with tandem mass spectrometry (LC/LC-MS/MS) for large-scale protein analysis: the yeast proteome. , 2003, Journal of proteome research.

[46]  R. Haslam,et al.  Molecular cloning, bacterial expression and properties of Rab31 and Rab32. , 2002, European journal of biochemistry.

[47]  長谷川俊史 Functional expression of the high affinity receptor for IgE(FcεRI)in human platelets and its intracellular expression in human megakaryocytes(ヒト血小板および巨核球における高親和性IgE受容体(FcεRI)の発現および機能の検討) , 1999 .

[48]  J. Seilhamer,et al.  A comparison of selected mRNA and protein abundances in human liver , 1997, Electrophoresis.

[49]  G. Stein,et al.  Changes in the stability of a human H3 histone mRNA during the HeLa cell cycle. , 1991, Molecular and cellular biology.