Last updated: 2019-06-14

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Knit directory: apaQTL/analysis/

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    Modified:   analysis/Readdistagainstfeatures.Rmd
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Rmd bf91cb3 brimittleman 2019-06-14 add location plot

In this analysis I want to look at the location of the apaQTLs first looking at distance to PAS. Until now I have been using the distance to the peak and have not flipped the strand. This showed me QTLs are close to the PAS but was not the most correct way to do this.

library(workflowr)
This is workflowr version 1.3.0
Run ?workflowr for help getting started
library(tidyverse)
── Attaching packages ────────────────────────────────────────────────── tidyverse 1.2.1 ──
✔ ggplot2 3.1.1       ✔ 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(cowplot)

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

    ggsave
PAS=read.table("../data/PAS/APAPAS_GeneLocAnno.5perc.bed",col.names = c("chr", "start", "PASloc", "name", "score", "strand"), stringsAsFactors = F )%>% separate(name, into=c("peakNum", "geneloc"), sep=":") %>% mutate(peak=paste("peak", peakNum, sep="")) %>% select(PASloc, peak)

Distance to PAS

Total:

totQTLs=read.table("../data/apaQTLs/Total_apaQTLs4pc_5fdr.WITHSTRAND.bed",stringsAsFactors = F, header=T)%>%
  separate(name, into=c("gene", "peak", "loc"), sep=":") %>% 
  inner_join(PAS, by="peak") %>% 
  mutate(distance=SNPend-PASloc, dist2PAS=ifelse(strand=="-", -1 *distance, distance))
ggplot(totQTLs, aes(x=dist2PAS, by=loc, fill=loc)) + geom_histogram(bins=100)

Plot by proportion:

ggplot(totQTLs, aes(x=dist2PAS, fill=loc)) + geom_histogram( bins=100) + facet_grid(~loc)

Nuclear

nucQTLs=read.table("../data/apaQTLs/Nuclear_apaQTLs4pc_5fdr.WITHSTRAND.bed",stringsAsFactors = F, header=T)%>%
  separate(name, into=c("gene", "peak", "loc"), sep=":") %>% 
  inner_join(PAS, by="peak") %>% 
  mutate(distance=SNPend-PASloc, dist2PAS=ifelse(strand=="-", -1 *distance, distance))
ggplot(nucQTLs, aes(x=dist2PAS, by=loc, fill=loc)) + geom_histogram(bins=100)

ggplot(nucQTLs, aes(x=dist2PAS, fill=loc)) + geom_histogram( bins=100) + facet_grid(~loc)

Metagene plot

I want to plot by normalized position in the gene.

genes=tss=read.table("../../genome_anotation_data/refseq.ProteinCoding.bed",col.names = c("chrom", "Genestart", "Geneend", "gene", "score", "strand") ,stringsAsFactors = F) %>% select(Genestart, Geneend, gene)

Total:

totQTLs_gene= totQTLs %>% inner_join(genes, by="gene")%>% mutate(geneLength=Geneend-Genestart) %>% mutate(dist2QTLnostrand= as.numeric(SNPend)-as.numeric(Genestart), dist2QTL=ifelse(strand=="-", -1 *dist2QTLnostrand, dist2QTLnostrand), propGene=dist2QTL/geneLength)  %>% filter(propGene>-5 & propGene<5)
ggplot(totQTLs_gene, aes(x=propGene, fill=loc)) + geom_histogram(bins=50)  + labs(x="Proportion of gene body", y="number QTLs", title="Total apaQTLs") + geom_vline(xintercept =0,color= "black") + geom_vline(xintercept =1,color= "black")

There are about 48 QTLs outside.

I can look at only those in the gene body:

totQTLs_gene_body= totQTLs_gene %>% filter(propGene>=0, propGene<=1)

ggplot(totQTLs_gene_body, aes(x=propGene, fill=loc)) + geom_histogram(bins=50)  + labs(x="Proportion of gene body", y="number QTLs", title="Total apaQTLs in gene body") 

There are 181 in the gene body

Nuclear:

nucQTLs_gene= nucQTLs %>% inner_join(genes, by="gene")%>% mutate(geneLength=Geneend-Genestart) %>% mutate(dist2QTLnostrand= as.numeric(SNPend)-as.numeric(Genestart), dist2QTL=ifelse(strand=="-", -1 *dist2QTLnostrand, dist2QTLnostrand), propGene=dist2QTL/geneLength) %>% filter(propGene>-5 & propGene<5)
ggplot(nucQTLs_gene, aes(x=propGene, fill=loc)) + geom_histogram(bins=50)  + labs(x="Proportion of gene body", y="number QTLs", title="Nuclear apaQTLs") + geom_vline(xintercept =0,color= "black") + geom_vline(xintercept =1,color= "black")

there are 77 outside of 500% of gene body

nucQTLs_gene_body= nucQTLs_gene %>% filter(propGene>=0, propGene<=1)

ggplot(nucQTLs_gene_body, aes(x=propGene, fill=loc)) + geom_histogram(bins=50)  + labs(x="Proportion of gene body", y="number QTLs", title="Nuclear apaQTLs in gene body") 

334 are in the gene body.


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   forcats_0.3.0   stringr_1.3.1   dplyr_0.8.0.1  
 [5] purrr_0.3.2     readr_1.3.1     tidyr_0.8.3     tibble_2.1.1   
 [9] ggplot2_3.1.1   tidyverse_1.2.1 workflowr_1.3.0

loaded via a namespace (and not attached):
 [1] Rcpp_1.0.0       cellranger_1.1.0 pillar_1.3.1     compiler_3.5.1  
 [5] git2r_0.25.2     plyr_1.8.4       tools_3.5.1      digest_0.6.18   
 [9] lubridate_1.7.4  jsonlite_1.6     evaluate_0.12    nlme_3.1-137    
[13] gtable_0.2.0     lattice_0.20-38  pkgconfig_2.0.2  rlang_0.3.1     
[17] cli_1.0.1        rstudioapi_0.10  yaml_2.2.0       haven_1.1.2     
[21] withr_2.1.2      xml2_1.2.0       httr_1.3.1       knitr_1.20      
[25] hms_0.4.2        generics_0.0.2   fs_1.2.6         rprojroot_1.3-2 
[29] grid_3.5.1       tidyselect_0.2.5 glue_1.3.0       R6_2.3.0        
[33] readxl_1.1.0     rmarkdown_1.10   reshape2_1.4.3   modelr_0.1.2    
[37] magrittr_1.5     whisker_0.3-2    backports_1.1.2  scales_1.0.0    
[41] htmltools_0.3.6  rvest_0.3.2      assertthat_0.2.0 colorspace_1.3-2
[45] labeling_0.3     stringi_1.2.4    lazyeval_0.2.1   munsell_0.5.0   
[49] broom_0.5.1      crayon_1.3.4