| Version: | 0.99.0 | 
| Date: | 2017-04-25 | 
| Title: | Kinase-Substrate Enrichment Analysis | 
| Description: | Infers relative kinase activity from phosphoproteomics data using the method described by Casado et al. (2013) <doi:10.1126/scisignal.2003573>. | 
| Author: | Danica D. Wiredja | 
| Maintainer: | Danica D. Wiredja <dwiredja@gmail.com> | 
| License: | MIT + file LICENSE | 
| Depends: | R (≥ 2.10) | 
| Imports: | gplots, graphics, stats, grDevices, utils | 
| Suggests: | knitr, rmarkdown | 
| VignetteBuilder: | knitr | 
| LazyData: | true | 
| RoxygenNote: | 6.0.1 | 
| NeedsCompilation: | no | 
| Packaged: | 2017-05-02 13:25:34 UTC; Danica | 
| Repository: | CRAN | 
| Date/Publication: | 2017-05-02 16:11:49 UTC | 
Kinase-Substrate (K-S) Relationship Dataset
Description
K-S annotations from PhosphoSitePlus and NetworKIN predictions; This is an abbreviated version of the full dataset used purely for demonstration; please go to the GitHub page for access to the complete file: github.com/casecpb/KSEA/
Usage
data(KSData)
Format
abbreviated dataframe containing the kinase-substrate annotations and source
References
Hornbeck et al. (2015) Nucleic Acids Res. 43:D512-20
Horn et al. (2014) Nature Methods 11(6):603-4
The KSEA App Analysis (KSEA Bar Plot Only)
Description
Takes a formatted phoshoproteomics data input and returns just the summary bar plot of kinase scores
Usage
KSEA.Barplot(KSData, PX, NetworKIN, NetworKIN.cutoff, m.cutoff, p.cutoff,
  export)
Arguments
| KSData | the Kinase-Substrate dataset uploaded from the file prefaced with "PSP&NetworKIN_" available from github.com/casecpb/KSEA/ | 
| PX | the experimental data file formatted as described in the KSEA.Complete() documentation | 
| NetworKIN | a binary input of TRUE or FALSE, indicating whether or not to include NetworKIN predictions; NetworKIN = TRUE means inclusion of NetworKIN predictions | 
| NetworKIN.cutoff | a numeric value between 1 and infinity setting the minimum NetworKIN score (can be left out if NetworKIN = FALSE) | 
| m.cutoff | a numeric value between 0 and infinity indicating the min. # of substrates a kinase must have to be included in the bar plot output | 
| p.cutoff | a numeric value between 0 and 1 indicating the p-value cutoff for indicating significant kinases in the bar plot | 
| export | a binary input of TRUE or FALSE, indicating whether or not to export the bar plot as a .tiff image into the working directory | 
Value
creates the bar plot output highlighting key kinase results
References
Casado et al. (2013) Sci Signal. 6(268):rs6
Hornbeck et al. (2015) Nucleic Acids Res. 43:D512-20
Horn et al. (2014) Nature Methods 11(6):603-4
Examples
KSEA.Barplot(KSData, PX, NetworKIN=TRUE, NetworKIN.cutoff=5, 
             m.cutoff=5, p.cutoff=0.01, export=FALSE)
KSEA.Barplot(KSData, PX, NetworKIN=TRUE, NetworKIN.cutoff=5, 
             m.cutoff=8, p.cutoff=0.05, export=TRUE)
KSEA.Barplot(KSData, PX, NetworKIN=FALSE, m.cutoff=2, p.cutoff=0.05, export=TRUE)
The Complete KSEA App Analysis
Description
Takes a formatted phoshoproteomics data input and performs KSEA calculations to infer relative kinase activities
Usage
KSEA.Complete(KSData, PX, NetworKIN, NetworKIN.cutoff, m.cutoff, p.cutoff)
Arguments
| KSData | the Kinase-Substrate dataset uploaded from the file prefaced with "PSP&NetworKIN_" available from github.com/casecpb/KSEA/ | 
| PX | the experimental data file formatted exactly as described below; must have 6 columns in the exact order: Protein, Gene, Peptide, Residue.Both, p, FC; cannot have NA values, or else the entire peptide row is deleted; Description of each column in PX: 
 | 
| NetworKIN | a binary input of TRUE or FALSE, indicating whether or not to include NetworKIN predictions; NetworKIN = TRUE means inclusion of NetworKIN predictions | 
| NetworKIN.cutoff | a numeric value between 1 and infinity setting the minimum NetworKIN score (can be left out if NetworKIN = FALSE) | 
| m.cutoff | a numeric value between 0 and infinity indicating the min. # of substrates a kinase must have to be included in the bar plot output | 
| p.cutoff | a numeric value between 0 and 1 indicating the p-value cutoff for indicating significant kinases in the bar plot | 
Value
creates the following outputs that are deposited into your working directory: a bar plot highlighting key kinase results, a .csv file of all KSEA kinase scores, and a .csv file listing all kinase-substrate relationships used for the calculations
References
Casado et al. (2013) Sci Signal. 6(268):rs6
Hornbeck et al. (2015) Nucleic Acids Res. 43:D512-20
Horn et al. (2014) Nature Methods 11(6):603-4
Examples
KSEA.Complete(KSData, PX, NetworKIN=TRUE, NetworKIN.cutoff=5, m.cutoff=5, p.cutoff=0.01)
KSEA.Complete(KSData, PX, NetworKIN=FALSE, m.cutoff=2, p.cutoff=0.05)
The KSEA App Analysis (KSEA Heatmap Only)
Description
Takes a list of the KSEA kinase score outputs from KSEA.Scores() and creates a merged heatmap (only applicable for multi-treatment studies)
Usage
KSEA.Heatmap(score.list, sample.labels, stats, m.cutoff, p.cutoff,
  sample.cluster)
Arguments
| score.list | the data frame outputs from the KSEA.Scores() function, compiled in a list format | 
| sample.labels | a character vector of all the sample names for heatmap annotation; the names must be in the same order as the data in score.list; please avoid long names, as they may get cropped in the final image | 
| stats | character string of either "p.value" or "FDR" indicating the data column to use for marking statistically significant scores | 
| m.cutoff | a numeric value between 0 and infinity indicating the min. # of substrates a kinase must have to be included in the heatmap | 
| p.cutoff | a numeric value between 0 and 1 indicating the p-value/FDR cutoff for indicating significant kinases in the heatmap | 
| sample.cluster | a binary input of TRUE or FALSE, indicating whether or not to perform hierarchical clustering of the sample columns | 
Value
exports a .png heatmap image highlighting the merged datasets; heatmap was generated using the heatmap.2() function (gplots package); asterisks mark scores that met the statistical cutoff, as defined by p.cutoff; blue color indicates negative kinase score, and red indicates positive kinase score
References
Casado et al. (2013) Sci Signal. 6(268):rs6
Hornbeck et al. (2015) Nucleic Acids Res. 43:D512-20
Horn et al. (2014) Nature Methods 11(6):603-4
Examples
#The score.list input must be a list of the data frame outputs from KSEA.Scores() function
#KSEA.Scores.1, KSEA.Scores.2, and KSEA.Scores.3 are all 
#sample datasets provided within this package
KSEA.Heatmap(score.list=list(KSEA.Scores.1, KSEA.Scores.2, KSEA.Scores.3), 
             sample.labels=c("Tumor.A", "Tumor.B", "Tumor.C"), 
             stats="p.value", m.cutoff=3, p.cutoff=0.05, sample.cluster=TRUE)
The KSEA App Analysis (K-S Dataset Only)
Description
Takes a formatted phoshoproteomics data input and returns just the kinase-subtrate (K-S) annotations used for KSEA calculations
Usage
KSEA.KS_table(KSData, PX, NetworKIN, NetworKIN.cutoff)
Arguments
| KSData | the Kinase-Substrate dataset uploaded from the file prefaced with "PSP&NetworKIN_" available from github.com/casecpb/KSEA/ | 
| PX | the experimental data file formatted as described in the KSEA.Complete() documentation | 
| NetworKIN | a binary input of TRUE or FALSE, indicating whether or not to include NetworKIN predictions; NetworKIN = TRUE means inclusion of NetworKIN predictions | 
| NetworKIN.cutoff | a numeric value between 1 and infinity setting the minimum NetworKIN score (can be left out if NetworKIN = FALSE) | 
Value
creates a new data frame in R with all kinase-substrate relationships used for the KSEA calculations
References
Casado et al. (2013) Sci Signal. 6(268):rs6
Hornbeck et al. (2015) Nucleic Acids Res. 43:D512-20
Horn et al. (2014) Nature Methods 11(6):603-4
Examples
KSData.dataset = KSEA.KS_table(KSData, PX, NetworKIN=TRUE, NetworKIN.cutoff=3)
KSData.dataset = KSEA.KS_table(KSData, PX, NetworKIN=FALSE)
The KSEA App Analysis (KSEA Kinase Scores Only)
Description
Takes a formatted phoshoproteomics data input and returns just the KSEA kinase scores and statistics
Usage
KSEA.Scores(KSData, PX, NetworKIN, NetworKIN.cutoff)
Arguments
| KSData | the Kinase-Substrate dataset uploaded from the file prefaced with "PSP&NetworKIN_" available from github.com/casecpb/KSEA/ | 
| PX | the experimental data file formatted as described in the KSEA.Complete() documentation | 
| NetworKIN | a binary input of TRUE or FALSE, indicating whether or not to include NetworKIN predictions; NetworKIN = TRUE means inclusion of NetworKIN predictions | 
| NetworKIN.cutoff | a numeric value between 1 and infinity setting the minimum NetworKIN score (can be left out if NetworKIN = FALSE) | 
Value
creates a new data frame in R with all the KSEA kinase scores, along with each one's statistical assessment
References
Casado et al. (2013) Sci Signal. 6(268):rs6
Hornbeck et al. (2015) Nucleic Acids Res. 43:D512-20
Horn et al. (2014) Nature Methods 11(6):603-4
Examples
scores = KSEA.Scores(KSData, PX, NetworKIN=TRUE, NetworKIN.cutoff=3)
scores = KSEA.Scores(KSData, PX, NetworKIN=FALSE)
One of the 3 datasets for heatmap plotting
Description
A sample KSEA.Scores output generated from the KSEA.Scores() function (or alternatively, the "KSEA Kinase Scores.csv" output from the KSEA.Complete() function, loaded into R)
Usage
data(KSEA.Scores.1)
Format
dataframe containing 7 columns in the exact order as listed below.
- "KinaseGene" the HUGO gene name of the kinase 
- "mS" the mean log2FC of all the kinase's identified substrates 
- "Enrichment" the enrichment score (refer to Casado et al. (2013) Sci. Signal., 6, rs6-rs6) 
- "m" the number of experimentally-identifed substrates annotating to that kinase 
- "z.score" the normalized kinase score 
- "p.value" the statistical assessment of the kinase score 
- "FDR" the p-value adjusted for multiple hypothesis testing by the Benjamin-Hochberg method 
References
unpublished data
One of the 3 datasets for heatmap plotting
Description
A sample KSEA.Scores output generated from the KSEA.Scores() function (or alternatively, the "KSEA Kinase Scores.csv" output from the KSEA.Complete() function, loaded into R)
Usage
data(KSEA.Scores.2)
Format
dataframe containing 7 columns in the exact order as listed below.
- "KinaseGene" the HUGO gene name of the kinase 
- "mS" the mean log2FC of all the kinase's identified substrates 
- "Enrichment" the enrichment score (refer to Casado et al. (2013) Sci. Signal., 6, rs6-rs6) 
- "m" the number of experimentally-identifed substrates annotating to that kinase 
- "z.score" the normalized kinase score 
- "p.value" the statistical assessment of the kinase score 
- "FDR" the p-value adjusted for multiple hypothesis testing by the Benjamin-Hochberg method 
References
unpublished data
One of the 3 datasets for heatmap plotting
Description
A sample KSEA.Scores output generated from the KSEA.Scores() function (or alternatively, the "KSEA Kinase Scores.csv" output from the KSEA.Complete() function, loaded into R)
Usage
data(KSEA.Scores.3)
Format
dataframe containing 7 columns in the exact order as listed below.
- "KinaseGene" the HUGO gene name of the kinase 
- "mS" the mean log2FC of all the kinase's identified substrates 
- "Enrichment" the enrichment score (refer to Casado et al. (2013) Sci. Signal., 6, rs6-rs6) 
- "m" the number of experimentally-identifed substrates annotating to that kinase 
- "z.score" the normalized kinase score 
- "p.value" the statistical assessment of the kinase score 
- "FDR" the p-value adjusted for multiple hypothesis testing by the Benjamin-Hochberg method 
References
unpublished data
PX dataset for KSEA calculations
Description
A sample PX dataset of the experimental phosphoproteomics input
Usage
data(PX)
Format
the experimental data file must be formatted exactly as described below; must have 6 columns in the exact order: Protein, Gene, Peptide, Residue.Both, p, FC; cannot have NA values, or else the entire peptide row is deleted; Description of each column in PX:
- "Protein" the Uniprot ID for the parent protein 
- "Gene" the HUGO gene name for the parent protein 
- "Peptide" the peptide sequence 
- "Residue.Both" all phosphosites from that peptide, separated by semicolons if applicable; must be formatted as the single amino acid abbrev. with the residue position (e.g. S102) 
- "p" the p-value of that peptide (if none calculated, please write "NULL", cannot be NA) 
- "FC" the fold change (not log-transformed); usually the control sample is the denominator 
References
unpublished data