PIIKA 2: An Expanded, Web-Based Platform for Analysis of Kinome Microarray Data

Kinome microarrays are comprised of peptides that act as phosphorylation targets for protein kinases. This platform is growing in popularity due to its ability to measure phosphorylation-mediated cellular signaling in a high-throughput manner. While software for analyzing data from DNA microarrays has also been used for kinome arrays, differences between the two technologies and associated biologies previously led us to develop Platform for Intelligent, Integrated Kinome Analysis (PIIKA), a software tool customized for the analysis of data from kinome arrays. Here, we report the development of PIIKA 2, a significantly improved version with new features and improvements in the areas of clustering, statistical analysis, and data visualization. Among other additions to the original PIIKA, PIIKA 2 now allows the user to: evaluate statistically how well groups of samples cluster together; identify sets of peptides that have consistent phosphorylation patterns among groups of samples; perform hierarchical clustering analysis with bootstrapping; view false negative probabilities and positive and negative predictive values for t-tests between pairs of samples; easily assess experimental reproducibility; and visualize the data using volcano plots, scatterplots, and interactive three-dimensional principal component analyses. Also new in PIIKA 2 is a web-based interface, which allows users unfamiliar with command-line tools to easily provide input and download the results. Collectively, the additions and improvements described here enhance both the breadth and depth of analyses available, simplify the user interface, and make the software an even more valuable tool for the analysis of kinome microarray data. Both the web-based and stand-alone versions of PIIKA 2 can be accessed via http://saphire.usask.ca.

[1]  E. Krebs,et al.  Role of multiple basic residues in determining the substrate specificity of cyclic AMP-dependent protein kinase. , 1977, The Journal of biological chemistry.

[2]  B. Trost,et al.  Divergent Immune Responses to Mycobacterium avium subsp. paratuberculosis Infection Correlate with Kinome Responses at the Site of Intestinal Infection , 2013, Infection and Immunity.

[3]  Jonathan M. Garibaldi,et al.  vrmlgen: An R Package for 3D Data Visualization on the Web , 2010 .

[4]  B. Trost,et al.  A Systematic Approach for Analysis of Peptide Array Kinome Data , 2012, Science Signaling.

[5]  X. Cui,et al.  Statistical tests for differential expression in cDNA microarray experiments , 2003, Genome Biology.

[6]  M. Mrksich,et al.  Peptide chips for the quantitative evaluation of protein kinase activity , 2002, Nature Biotechnology.

[7]  R Core Team,et al.  R: A language and environment for statistical computing. , 2014 .

[8]  Hidetoshi Shimodaira,et al.  Approximately unbiased tests of regions using multistep-multiscale bootstrap resampling , 2004, math/0508602.

[9]  E. Humble,et al.  The minimum substrate of cyclic AMP-stimulated protein kinase, as studied by synthetic peptides representing the phosphorylatable site of pyruvate kinase (type L) of rat liver. , 1976, Biochemical and biophysical research communications.

[10]  Jeffrey W Pollard,et al.  Gene Expression Analysis of Macrophages That Facilitate Tumor Invasion Supports a Role for Wnt-Signaling in Mediating Their Activity in Primary Mammary Tumors , 2009, The Journal of Immunology.

[11]  Frank Buttgereit,et al.  Rapid immunosuppressive effects of glucocorticoids mediated through Lck and Fyn. , 2005, Blood.

[12]  A. Potter,et al.  Altered Toll-Like Receptor 9 Signaling in Mycobacterium avium subsp. paratuberculosis-Infected Bovine Monocytes Reveals Potential Therapeutic Targets , 2012, Infection and Immunity.

[13]  James A. Thomson,et al.  Induced pluripotent stem cells from a spinal muscular atrophy patient , 2009, Nature.

[14]  Greg Finak,et al.  Gene expression signatures of morphologically normal breast tissue identify basal-like tumors , 2006, Breast Cancer Research.

[15]  J. Felsenstein CONFIDENCE LIMITS ON PHYLOGENIES: AN APPROACH USING THE BOOTSTRAP , 1985, Evolution; international journal of organic evolution.

[16]  P. Jahrling,et al.  Systems Kinomics Demonstrates Congo Basin Monkeypox Virus Infection Selectively Modulates Host Cell Signaling Responses as Compared to West African Monkeypox Virus , 2011, Molecular & Cellular Proteomics.

[17]  J. Graber,et al.  Global changes in processing of mRNA 3' untranslated regions characterize clinically distinct cancer subtypes. , 2009, Cancer research.

[18]  C. Pieterse,et al.  Are Small GTPases Signal Hubs in Sugar-Mediated Induction of Fructan Biosynthesis? , 2009, PloS one.

[19]  M. Peppelenbosch,et al.  Major remodelling of the murine stem cell kinome following differentiation in the hematopoietic compartment. , 2011, Journal of proteome research.

[20]  A. Kusalik,et al.  Mycobacterium avium subsp. paratuberculosis Inhibits Gamma Interferon-Induced Signaling in Bovine Monocytes: Insights into the Cellular Mechanisms of Johne's Disease , 2012, Infection and Immunity.

[21]  M. Peppelenbosch Kinome Profiling , 2012, Scientifica.

[22]  M. Mrksich,et al.  Towards quantitative assays with peptide chips: a surface engineering approach. , 2002, Trends in biotechnology.

[23]  Hidetoshi Shimodaira An approximately unbiased test of phylogenetic tree selection. , 2002, Systematic biology.

[24]  Hidetoshi Shimodaira,et al.  Pvclust: an R package for assessing the uncertainty in hierarchical clustering , 2006, Bioinform..

[25]  Matthew R. Laird,et al.  Protein Protein Interaction Network Evaluation for Identifying Potential Drug Targets , 2009 .

[26]  A. Regev,et al.  An embryonic stem cell–like gene expression signature in poorly differentiated aggressive human tumors , 2008, Nature Genetics.

[27]  T. van Wezel,et al.  Kinome profiling of chondrosarcoma reveals SRC-pathway activity and dasatinib as option for treatment. , 2009, Cancer research.

[28]  T. Ritsema,et al.  Kinome profiling of sugar signaling in plants using multiple platforms , 2009, Plant signaling & behavior.

[29]  Scott Napper,et al.  Genome to Kinome: Species-Specific Peptide Arrays for Kinome Analysis , 2009, Science Signaling.

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