Proportion eQTLs explained

Last updated: 2019-06-12

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Knit directory: apaQTL/analysis/

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Unstaged changes:
    Modified:   analysis/DiffIsoAnalysis.Rmd
    Modified:   analysis/PASusageQC.Rmd
    Modified:   analysis/Readdistagainstfeatures.Rmd
    Modified:   analysis/choosePCs.Rmd
    Modified:   analysis/corrbetweenind.Rmd
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    Modified:   analysis/nucintronicanalysis.Rmd
    Modified:   analysis/overlapapaqtlsandeqtls.Rmd
    Modified:   analysis/rna_netseq_h3k12ac.Rmd
    Modified:   code/BothFracDTPlotGeneRegions.sh
    Modified:   code/Snakefile
    Deleted:    code/Upstream10Bases_general.py
    Modified:   code/apaQTLCorrectPvalMakeQQ.R
    Modified:   code/apaQTL_permuted.sh
    Modified:   code/apaQTLsnake.err
    Modified:   code/bam2bw.sh
    Modified:   code/bed2saf.py
    Modified:   code/cluster.json
    Modified:   code/config.yaml
    Deleted:    code/test.txt

Note that any generated files, e.g. HTML, png, CSS, etc., are not included in this status report because it is ok for generated content to have uncommitted changes.

These are the previous versions of the R Markdown and HTML files. If you’ve configured a remote Git repository (see ?wflow_git_remote), click on the hyperlinks in the table below to view them.

File	Version	Author	Date	Message
Rmd	178c5dc	brimittleman	2019-06-12	new geno
html	6b164c8	brimittleman	2019-06-07	Build site.
Rmd	b39620d	brimittleman	2019-06-07	add bonfor results
html	458e494	brimittleman	2019-06-07	Build site.
Rmd	32091ee	brimittleman	2019-06-07	more prop explained to new analysis

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(workflowr)

This is workflowr version 1.3.0
Run ?workflowr for help getting started

library(reshape2)


Attaching package: 'reshape2'

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

    smiths

I need to fix the explained_FDR10.sort.txt and unexplained_FDR10.sort.txt files because right now this file has multiple genes per snp.

python fixExandUnexeQTL.py ../data/Li_eQTLs/explained_FDR10.sort.txt ../data/Li_eQTLs/explained_FDR10.sort_FIXED.txt
python fixExandUnexeQTL.py ../data/Li_eQTLs/unexplained_FDR10.sort.txt ../data/Li_eQTLs/unexplained_FDR10.sort_FIXED.txt

There are 1195 explained and 814 unexplained eQTLs. I will next look at each of these in my apadata.

Convert nominal results to have snps rather than rsids:

python convertNominal2SNPLOC.py Total
python convertNominal2SNPLOC.py Nuclear

mkdir ../data/overlapeQTL_try2
sbatch run_getapafromeQTL.sh

total

I can group the unexplained by gene and snp then I can ask if there is at least 1 significat peak for each of these.

I will use the bonforoni correction here and multiply the pvalue by the number of peaks in the gene:snp association.

nomnames=c("peakID", 'snp','dist', 'pval', 'slope')
totalapaUnexplained=read.table("../data/overlapeQTL_try2/apaTotal_unexplainedQTLs.txt", stringsAsFactors = F, col.names = nomnames) %>% separate(peakID, into=c("chr","start","end","geneID"), sep=":") %>% separate(geneID, into=c("gene", "loc", "strand", "PASnum"), sep="_") %>% group_by(gene, snp)  %>% mutate(nPeaks=n(), adjPval=pval* nPeaks) %>%  slice(which.min(adjPval))

totalapaUnexplained_sig= totalapaUnexplained %>% filter(adjPval<.05)

Look at distribution of these pvals:

ggplot(totalapaUnexplained, aes(x=adjPval)) + geom_histogram(bins=50)

Proportion explained:

nrow(totalapaUnexplained_sig)/nrow(totalapaUnexplained)

[1] 0.1287879

I tested 528 unexplained eQTLs in the total fraction and 68 have a bonforoni corrected significant peak.

Compare to explained eQTLS:

totalapaexplained=read.table("../data/overlapeQTL_try2/apaTotal_explainedQTLs.txt", stringsAsFactors = F, col.names = nomnames) %>% separate(peakID, into=c("chr","start","end","geneID"), sep=":") %>% separate(geneID, into=c("gene", "loc", "strand", "PASnum"), sep="_") %>% group_by(gene, snp) %>%  mutate(nPeaks=n(), adjPval=pval* nPeaks) %>%  slice(which.min(adjPval))

totalapaexplained_sig= totalapaexplained %>% filter(adjPval<.05)

nrow(totalapaexplained_sig)/nrow(totalapaexplained)

[1] 0.1157601

I am testing 717 explained eQTLs and of those 83 have a bonforoni corrected significant peak.

difference of proportions:

prop.test(x=c(nrow(totalapaUnexplained_sig),nrow(totalapaexplained_sig)), n=c(nrow(totalapaUnexplained),nrow(totalapaexplained)))


    2-sample test for equality of proportions with continuity
    correction

data:  c(nrow(totalapaUnexplained_sig), nrow(totalapaexplained_sig)) out of c(nrow(totalapaUnexplained), nrow(totalapaexplained))
X-squared = 0.36972, df = 1, p-value = 0.5432
alternative hypothesis: two.sided
95 percent confidence interval:
 -0.02555884  0.05161437
sample estimates:
   prop 1    prop 2 
0.1287879 0.1157601

ggplot(totalapaUnexplained_sig,aes(x=loc)) + geom_histogram(stat="count",aes(y=..count../sum(..count..))) + labs(y="Proportion", title = "Total apaQTLs explaining eQTLs")

Warning: Ignoring unknown parameters: binwidth, bins, pad

totalapaUnexplained_sig_loc= totalapaUnexplained_sig %>% group_by(loc) %>% summarise(nLocTotalUn=n()) %>% mutate(propTotalUn=nLocTotalUn/nrow(totalapaUnexplained_sig))
totalapaexplained_sig_loc= totalapaexplained_sig %>% group_by(loc) %>% summarise(nLocTotalEx=n()) %>% mutate(propTotalEx=nLocTotalEx/nrow(totalapaexplained_sig))

BothTotalLoc=totalapaUnexplained_sig_loc %>% full_join(totalapaexplained_sig_loc,by="loc") %>%  replace_na(list(propTotalUn = 0, nLocTotalUn = 0,propTotalEx=0,nLocTotalEx=0  ))

BothTotalLoc

# A tibble: 5 x 5
  loc    nLocTotalUn propTotalUn nLocTotalEx propTotalEx
  <chr>        <dbl>       <dbl>       <dbl>       <dbl>
1 end              6      0.0882           9      0.108 
2 intron          24      0.353           27      0.325 
3 utr3            37      0.544           44      0.530 
4 utr5             1      0.0147           1      0.0120
5 cds              0      0                2      0.0241

nuclear

nuclearapaUnexplained=read.table("../data/overlapeQTL_try2/apaNuclear_unexplainedQTLs.txt", stringsAsFactors = F, col.names = nomnames) %>% separate(peakID, into=c("chr","start","end","geneID"), sep=":") %>% separate(geneID, into=c("gene", "loc", "strand", "PASnum"), sep="_") %>% group_by(gene, snp)  %>%  mutate(nPeaks=n(), adjPval=pval* nPeaks) %>%  slice(which.min(adjPval))

nuclearapaUnexplained_sig= nuclearapaUnexplained %>% filter(adjPval<.05)

nrow(nuclearapaUnexplained_sig)/nrow(nuclearapaUnexplained)

[1] 0.1660448

I tested 536 unexplained eQTLs in the nuclear fraction and 89 have a bonforoni corrected significant peak.

nuclearapaexplained=read.table("../data/overlapeQTL_try2/apaNuclear_explainedQTLs.txt", stringsAsFactors = F, col.names = nomnames) %>% separate(peakID, into=c("chr","start","end","geneID"), sep=":") %>% separate(geneID, into=c("gene", "loc", "strand", "PASnum"), sep="_") %>% group_by(gene, snp) %>%  mutate(nPeaks=n(), adjPval=pval* nPeaks) %>%  slice(which.min(adjPval))

nuclearapaexplained_sig= nuclearapaexplained %>% filter(adjPval<.05)

nrow(nuclearapaexplained_sig)/nrow(nuclearapaexplained)

[1] 0.1241379

I tested 725 explained eQTLs in the nuclear fraction and 90 have a nominally significant peak. difference of proportions:

prop.test(x=c(nrow(nuclearapaUnexplained_sig),nrow(nuclearapaexplained_sig)), n=c(nrow(nuclearapaUnexplained),nrow(nuclearapaexplained)))


    2-sample test for equality of proportions with continuity
    correction

data:  c(nrow(nuclearapaUnexplained_sig), nrow(nuclearapaexplained_sig)) out of c(nrow(nuclearapaUnexplained), nrow(nuclearapaexplained))
X-squared = 4.1059, df = 1, p-value = 0.04273
alternative hypothesis: two.sided
95 percent confidence interval:
 0.0006796895 0.0831340006
sample estimates:
   prop 1    prop 2 
0.1660448 0.1241379

Nuclear are more likely to explain unexplained than expalined.

ggplot(nuclearapaUnexplained_sig,aes(x=loc))  + geom_histogram(stat="count",aes(y=..count../sum(..count..))) + labs(title = "Nuclear apaQTLs explaining eQTLs", y="Proportion")

Warning: Ignoring unknown parameters: binwidth, bins, pad

nuclearapaUnexplained_sig_loc= nuclearapaUnexplained_sig %>% group_by(loc) %>% summarise(nLocnuclearUn=n()) %>% mutate(propnuclearUn=nLocnuclearUn/nrow(nuclearapaUnexplained_sig))
nuclearapaexplained_sig_loc= nuclearapaexplained_sig %>% group_by(loc) %>% summarise(nLocnuclearEx=n()) %>% mutate(propnuclearEx=nLocnuclearEx/nrow(nuclearapaexplained_sig))

BothnuclearLoc=nuclearapaUnexplained_sig_loc %>% full_join(nuclearapaexplained_sig_loc,by="loc") %>%  replace_na(list(propnuclearUn = 0, nLocnuclearUn = 0,propnuclearEx=0,nLocnuclearEx=0  ))

BothnuclearLoc

# A tibble: 5 x 5
  loc    nLocnuclearUn propnuclearUn nLocnuclearEx propnuclearEx
  <chr>          <dbl>         <dbl>         <dbl>         <dbl>
1 cds                2        0.0225             4        0.0444
2 end                3        0.0337             5        0.0556
3 intron            39        0.438             48        0.533 
4 utr3              45        0.506             31        0.344 
5 utr5               0        0                  2        0.0222

prop.test(x=c(39,48), n=c(nrow(nuclearapaUnexplained_sig),nrow(nuclearapaexplained_sig)))


    2-sample test for equality of proportions with continuity
    correction

data:  c(39, 48) out of c(nrow(nuclearapaUnexplained_sig), nrow(nuclearapaexplained_sig))
X-squared = 1.2627, df = 1, p-value = 0.2611
alternative hypothesis: two.sided
95 percent confidence interval:
 -0.25207523  0.06181306
sample estimates:
   prop 1    prop 2 
0.4382022 0.5333333

prop.test(x=c(45,31), n=c(nrow(nuclearapaUnexplained_sig),nrow(nuclearapaexplained_sig)))


    2-sample test for equality of proportions with continuity
    correction

data:  c(45, 31) out of c(nrow(nuclearapaUnexplained_sig), nrow(nuclearapaexplained_sig))
X-squared = 4.1211, df = 1, p-value = 0.04235
alternative hypothesis: two.sided
95 percent confidence interval:
 0.007076415 0.315270651
sample estimates:
   prop 1    prop 2 
0.5056180 0.3444444

Unexplained are more likely to be in the 3’ UTR.

total v nuclear

prop.test(x=c(nrow(nuclearapaUnexplained_sig),nrow(totalapaUnexplained_sig)), n=c(nrow(nuclearapaUnexplained),nrow(totalapaUnexplained)))


    2-sample test for equality of proportions with continuity
    correction

data:  c(nrow(nuclearapaUnexplained_sig), nrow(totalapaUnexplained_sig)) out of c(nrow(nuclearapaUnexplained), nrow(totalapaUnexplained))
X-squared = 2.6465, df = 1, p-value = 0.1038
alternative hypothesis: two.sided
95 percent confidence interval:
 -0.007152311  0.081666106
sample estimates:
   prop 1    prop 2 
0.1660448 0.1287879

Differences in proportion by location

allLocProp=BothnuclearLoc %>% full_join(BothTotalLoc, by="loc") %>% select(loc,propnuclearUn,propnuclearEx,propTotalUn,propTotalEx )

allLocPropmelt= melt(allLocProp, id.vars = "loc") %>% mutate(Fraction=ifelse(grepl("Total", variable), "Total", "Nuclear"),eQTL=ifelse(grepl("Un", variable), "Unexplained", "Explained"))


ggplot(allLocPropmelt,aes(x=loc, fill=eQTL, y=value)) + geom_histogram(stat="identity", position = "dodge") + facet_grid(~Fraction)+ labs(y="Proportion of PAS", title="apaQTLs overlaping eQTLs by PAS location")

Warning: Ignoring unknown parameters: binwidth, bins, pad

This is a very stringent test. A less stringent way to get an upper bound would be to make an informed decision about which peak to use. This will make it so I am only testing one PAS per gene.

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] reshape2_1.4.3  workflowr_1.3.0 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_3.1.1   tidyverse_1.2.1

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] fansi_0.4.0      readxl_1.1.0     rmarkdown_1.10   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     utf8_1.1.4       stringi_1.2.4    lazyeval_0.2.1  
[49] munsell_0.5.0    broom_0.5.1      crayon_1.3.4

Proportion eQTLs explained

Briana Mittleman

6/7/2019

total

nuclear

total v nuclear