Last updated: 2019-04-24

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In this analysis I will create discriptive plots for the PAS identified in the 54 LCLs.

library(workflowr)
This is workflowr version 1.3.0
Run ?workflowr for help getting started
library(tidyverse)
── Attaching packages ──────────────────────────────────────────── tidyverse 1.2.1 ──
✔ ggplot2 3.1.0       ✔ purrr   0.3.2  
✔ tibble  2.1.1       ✔ dplyr   0.8.0.1
✔ tidyr   0.8.3       ✔ stringr 1.3.1  
✔ readr   1.3.1       ✔ forcats 0.3.0  
── Conflicts ─────────────────────────────────────────────── tidyverse_conflicts() ──
✖ dplyr::filter() masks stats::filter()
✖ dplyr::lag()    masks stats::lag()
library(reshape2)

Attaching package: 'reshape2'
The following object is masked from 'package:tidyr':

    smiths
library(cowplot)

Attaching package: 'cowplot'
The following object is masked from 'package:ggplot2':

    ggsave

Peaks per gene:

I want to plot how many genes have 0, 1, 2 and more than 2 PAS in the set. I need to join my PAS with the annotation to find out how many genes have 0 PAS.

pas=read.table("../data/PAS/APAPAS_GeneLocAnno.5perc.bed", header = F, stringsAsFactors = F, col.names = c("Chr", "start", "end", "PeakID", "score", "strand")) %>% separate(PeakID, into=c("peaknum", "geneAnno"), sep=":") %>% separate(geneAnno, into=c("Gene", "Loc"),sep="_")

pasbygene= pas %>% group_by(Gene) %>% summarise(PAS=n())

annotation=read.table("../../genome_anotation_data/RefSeq_annotations/ncbiRefSeq_FormatedallAnnotation.sort.bed", col.names = c("chr", "start", "end", "anno", "score", "strand")) %>% separate(anno, into=c("Loc", "Gene"),sep=":") %>% group_by(Gene) %>% summarise(annos=n()) %>% select(Gene)

PASallgene=annotation %>% full_join(pasbygene, by="Gene") %>% replace_na(list(PAS=0)) 

#group with 0,1,2,more than 2
PASallgene_grouped=PASallgene %>% mutate(Zero=ifelse(PAS==0,1, 0), One=ifelse(PAS==1,1,0), Multiple=ifelse(PAS>1,1,0))

Plot this:

Genes=c(sum(PASallgene_grouped$Zero),sum(PASallgene_grouped$One),sum(PASallgene_grouped$Multiple))
PAS=c("Zero", "One", "Multiple")
AllPAS=c(sum(PASallgene_grouped$Zero),sum(PASallgene_grouped$One),sum(PASallgene_grouped$Multiple))
GenebyPAS=as.data.frame(cbind(PAS,AllPAS))
GenebyPAS$AllPAS=as.numeric(as.character(GenebyPAS$AllPAS))

allPASplot=ggplot(GenebyPAS, aes(x="",y=AllPAS, fill=PAS)) + geom_bar(stat="identity", width=.5) + scale_fill_brewer(palette="GnBu") + labs(title="Identified PAS per Gene", y="Genes",x="All Indentified PAS")
allPASplot

Version Author Date
012892d brimittleman 2019-04-24
1fb7086 brimittleman 2019-04-23 
ggsave(allPASplot, file="../output/GeneswithAPApotentialAllPAS.png", width=3, height=5)

Subset and get stats for UTR

pasUTR=pas %>% filter(Loc=="utr3") %>% group_by(Gene) %>% summarise(PAS=n())

pasUTR_allgene=annotation %>% full_join(pasUTR, by="Gene") %>% replace_na(list(PAS=0)) 

PASUTRallgene_grouped=pasUTR_allgene %>% mutate(Zero=ifelse(PAS==0,1, 0), One=ifelse(PAS==1,1,0), Multiple=ifelse(PAS>1,1,0))


GenesUTR=c(sum(PASUTRallgene_grouped$Zero),sum(PASUTRallgene_grouped$One),sum(PASUTRallgene_grouped$Multiple))
UTR=c(sum(PASUTRallgene_grouped$Zero),sum(PASUTRallgene_grouped$One),sum(PASUTRallgene_grouped$Multiple))
GenebyPASUTR=as.data.frame(cbind(PAS,UTR))
GenebyPASUTR$UTR=as.numeric(as.character(GenebyPASUTR$UTR))

ggplot(GenebyPASUTR, aes(x="",y=UTR, fill=PAS)) + geom_bar(stat="identity")

Version Author Date
012892d brimittleman 2019-04-24
1fb7086 brimittleman 2019-04-23

Subset and get stats for Intron

pasIntron=pas %>% filter(Loc=="intron" | Loc=='utr3') %>% group_by(Gene) %>% summarise(PAS=n())

pasIntron_allgene=annotation %>% full_join(pasIntron, by="Gene") %>% replace_na(list(PAS=0)) 

pasIntronallgene_grouped=pasIntron_allgene %>% mutate(Zero=ifelse(PAS==0,1, 0), One=ifelse(PAS==1,1,0), Multiple=ifelse(PAS>1,1,0))


UTRandIntron=c(sum(pasIntronallgene_grouped$Zero),sum(pasIntronallgene_grouped$One),sum(pasIntronallgene_grouped$Multiple))
GenebyPASIntron=as.data.frame(cbind(PAS,UTRandIntron))
GenebyPASIntron$UTRandIntron=as.numeric(as.character(GenebyPASIntron$UTRandIntron))

ggplot(GenebyPASIntron, aes(x="",y=UTRandIntron, fill=PAS)) + geom_bar(stat="identity")

Version Author Date
012892d brimittleman 2019-04-24
1fb7086 brimittleman 2019-04-23

Make these side by side:

GenebyPASUTR_melt=melt(GenebyPASUTR, id.vars = "PAS", value.name = "Genes", variable.name = "Set")

GenebyPAS_melt=melt(GenebyPAS, id.vars = "PAS", value.name = "Genes", variable.name = "Set")

GenebyPASIntron_melt=melt(GenebyPASIntron, id.vars = "PAS", value.name = "Genes", variable.name = "Set")

GenebyPAStoplot=rbind(GenebyPAS_melt,GenebyPASUTR_melt,GenebyPASIntron_melt)

geneswithAPA=ggplot(GenebyPAStoplot, aes(x=Set,y=Genes, fill=PAS, by=Set)) + geom_bar(stat="identity")+ scale_fill_brewer(palette="YlGnBu") + labs(title="Genes with APA poential")

geneswithAPA

Version Author Date
012892d brimittleman 2019-04-24
1fb7086 brimittleman 2019-04-23 
ggsave(geneswithAPA, file="../output/GeneswithAPApotential.png")
Saving 7 x 5 in image
GenebyPAStoplot
       PAS          Set Genes
1     Zero       AllPAS 19518
2      One       AllPAS  2062
3 Multiple       AllPAS  5535
4     Zero          UTR 20539
5      One          UTR  3659
6 Multiple          UTR  2917
7     Zero UTRandIntron 19960
8      One UTRandIntron  2285
9 Multiple UTRandIntron  4870

Location of PAS

PAS_loc=pas %>% group_by(Loc) %>% summarise(nPAS=n())
loclabel=c("Coding", "Downstream", "Intronic", "3' UTR", "5' UTR")
PASLocPlot=ggplot(PAS_loc, aes(x=Loc, y=nPAS, fill=Loc)) + geom_bar(stat="identity",width=.5)+ scale_fill_brewer(palette = "YlGnBu") + labs(x="Gene location", y="Number of identified PAS", title="Location distribution for identified PAS") + theme(legend.position = "none")+ scale_x_discrete(labels= loclabel)+theme(axis.text.x = element_text(angle = 90, hjust = 1))
PASLocPlot

Version Author Date
012892d brimittleman 2019-04-24 
ggsave(PASLocPlot, file="../output/PASlocation.png")
Saving 7 x 5 in image

sessionInfo()
R version 3.5.1 (2018-07-02)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Scientific Linux 7.4 (Nitrogen)

Matrix products: default
BLAS/LAPACK: /software/openblas-0.2.19-el7-x86_64/lib/libopenblas_haswellp-r0.2.19.so

locale:
 [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C              
 [3] LC_TIME=en_US.UTF-8        LC_COLLATE=en_US.UTF-8    
 [5] LC_MONETARY=en_US.UTF-8    LC_MESSAGES=en_US.UTF-8   
 [7] LC_PAPER=en_US.UTF-8       LC_NAME=C                 
 [9] LC_ADDRESS=C               LC_TELEPHONE=C            
[11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C       

attached base packages:
[1] stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
 [1] cowplot_0.9.4   reshape2_1.4.3  forcats_0.3.0   stringr_1.3.1  
 [5] dplyr_0.8.0.1   purrr_0.3.2     readr_1.3.1     tidyr_0.8.3    
 [9] tibble_2.1.1    ggplot2_3.1.0   tidyverse_1.2.1 workflowr_1.3.0

loaded via a namespace (and not attached):
 [1] Rcpp_1.0.0         RColorBrewer_1.1-2 cellranger_1.1.0  
 [4] pillar_1.3.1       compiler_3.5.1     git2r_0.23.0      
 [7] plyr_1.8.4         tools_3.5.1        digest_0.6.18     
[10] lubridate_1.7.4    jsonlite_1.6       evaluate_0.12     
[13] nlme_3.1-137       gtable_0.2.0       lattice_0.20-38   
[16] pkgconfig_2.0.2    rlang_0.3.1        cli_1.0.1         
[19] rstudioapi_0.10    yaml_2.2.0         haven_1.1.2       
[22] withr_2.1.2        xml2_1.2.0         httr_1.3.1        
[25] knitr_1.20         hms_0.4.2          generics_0.0.2    
[28] fs_1.2.6           rprojroot_1.3-2    grid_3.5.1        
[31] tidyselect_0.2.5   glue_1.3.0         R6_2.3.0          
[34] readxl_1.1.0       rmarkdown_1.10     modelr_0.1.2      
[37] magrittr_1.5       whisker_0.3-2      backports_1.1.2   
[40] scales_1.0.0       htmltools_0.3.6    rvest_0.3.2       
[43] assertthat_0.2.0   colorspace_1.3-2   labeling_0.3      
[46] stringi_1.2.4      lazyeval_0.2.1     munsell_0.5.0     
[49] broom_0.5.1        crayon_1.3.4