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"cd", - "$INIT_CWD", - "&&", - "mkdir", - "-p", - "fire-test-data", - "&&", - "aws", - "s3", - "--no-sign-request", - "sync", - "--endpoint-url", - "https://s3.kopah.orci.washington.edu", - "s3://stergachis/public/FIRE/test-data", - "fire-test-data/", + "cd", + "$INIT_CWD", + "&&", + "mkdir", + "-p", + "fire-test-data", + "&&", + "rclone", + "sync", + ":s3,env_auth=false,provider=Other,endpoint=s3.kopah.orci.washington.edu:stergachis/public/FIRE/test-data", + "fire-test-data/", ] } test = { cmd = [ - "cd", - "$INIT_CWD/fire-test-data", - "&&", - "snakemake", - "-s", - "$PIXI_PROJECT_ROOT/workflow/Snakefile", - "--configfile", - "test.yaml", - "-k", + "cd", + "$INIT_CWD/fire-test-data", + "&&", + "snakemake", + "-s", + "$PIXI_PROJECT_ROOT/workflow/Snakefile", + "--configfile", + "test.yaml", + "-k", ], depends-on = [ - "test-data", + "test-data", ], clean-env = true } fire = { cmd = [ - "cd", - "$INIT_CWD", - "&&", - "snakemake", - "-s", - "$PIXI_PROJECT_ROOT/workflow/Snakefile", + "cd", + "$INIT_CWD", + "&&", + "snakemake", + "-s", + "$PIXI_PROJECT_ROOT/workflow/Snakefile", ] } slurm = { cmd = [ - "cd", - "$INIT_CWD", - "&&", - "snakemake", - "-s", - "$PIXI_PROJECT_ROOT/workflow/Snakefile", - "--profile", - "$PIXI_PROJECT_ROOT/profiles/slurm-executor", + "cd", + "$INIT_CWD", + "&&", + "snakemake", + "-s", + "$PIXI_PROJECT_ROOT/workflow/Snakefile", + "--profile", + "$PIXI_PROJECT_ROOT/profiles/slurm-executor", ] } [dependencies] @@ -63,7 +59,7 @@ snakemake = "==8.21" snakemake-executor-plugin-slurm = ">=0.11.2" snakefmt = "*" ruff = "*" -awscli = "2.22.*" +rclone = "*" taplo = "*" [pypi-dependencies] diff --git a/workflow/Snakefile b/workflow/Snakefile index f36e0da1c..dc1e50fdb 100644 --- a/workflow/Snakefile +++ b/workflow/Snakefile @@ -23,7 +23,7 @@ FULL_VERSION = f"v{os.environ.get("PIXI_PROJECT_VERSION", "UNK")}" # trim the FULL_VERSION to just the just the major and minor version VERSION = ".".join(FULL_VERSION.split(".")[:2]) -# use the full version if requested +# use the full version if requested if config.get("full-version", False): VERSION = FULL_VERSION @@ -130,11 +130,8 @@ rule all: # coverage information expand(rules.coverage.output, sm=MANIFEST.index, v=VERSION), expand(rules.exclude_from_shuffle.output, sm=MANIFEST.index, v=VERSION), - expand(rules.unreliable_coverage_regions.output, sm=MANIFEST.index, v=VERSION), expand( - rules.pileup.output.bed, - sm=MANIFEST.index, - v=VERSION, + rules.unreliable_coverage_regions.output.bed, sm=MANIFEST.index, v=VERSION ), # model results expand(rules.fire.output.cram, sm=MANIFEST.index, v=VERSION), diff --git a/workflow/rules/coverages.smk b/workflow/rules/coverages.smk index c354c76cc..9a0aa1394 100644 --- a/workflow/rules/coverages.smk +++ b/workflow/rules/coverages.smk @@ -17,7 +17,7 @@ rule genome_bedgraph: DEFAULT_ENV shell: """ - mosdepth -f {input.ref} -t {threads} tmp {input.cram} + mosdepth -F 4 -f {input.ref} -t {threads} tmp {input.cram} bgzip -cd tmp.per-base.bed.gz \ | LC_ALL=C sort --parallel={threads} -k1,1 -k2,2n -k3,3n -k4,4 \ | bgzip -@ {threads} \ diff --git a/workflow/rules/fire-peaks.smk b/workflow/rules/fire-peaks.smk index 07d15e0f6..dbaf99c31 100644 --- a/workflow/rules/fire-peaks.smk +++ b/workflow/rules/fire-peaks.smk @@ -76,10 +76,12 @@ rule fdr_table: "../envs/python.yaml" params: script=workflow.source_path("../scripts/fdr-table.py"), + threads: 16 resources: mem_mb=get_mem_mb_xl, shell: """ + export POLARS_MAX_THREADS={threads} MIN=$(cat {input.minimum}) MAX=$(cat {input.maximum}) python {params.script} -v 1 {input.shuffled} {output.tbl} --max-cov $MAX --min-cov $MIN @@ -124,6 +126,7 @@ rule fdr_track_chromosome: mem_mb=get_mem_mb_xl, shell: """ + export POLARS_MAX_THREADS={threads} python {params.script} -v 1 \ --fdr-table {input.fdr_tbl} \ {input.pileup} {output.bed} @@ -141,7 +144,7 @@ rule pileup: fofn=temp("temp/{sm}/fire/fire-{v}-pileup.fofn"), bed="results/{sm}/{sm}-fire-{v}-pileup.bed.gz", tbi="results/{sm}/{sm}-fire-{v}-pileup.bed.gz.tbi", - threads: 4 + threads: 8 conda: DEFAULT_ENV shell: @@ -231,6 +234,7 @@ rule fdr_peaks_by_fire_elements_chromosome: min_frac_accessible=MIN_FRAC_ACCESSIBLE, shell: """ + export POLARS_MAX_THREADS={threads} bgzip -cd {input.bed} \ | python {params.script} -v 1 \ --max-cov $(cat {input.maximum}) \ diff --git a/workflow/scripts/cov.py b/workflow/scripts/cov.py index 0754ef914..53044ec13 100644 --- a/workflow/scripts/cov.py +++ b/workflow/scripts/cov.py @@ -1,3 +1,7 @@ +import os + +os.environ["POLARS_MAX_THREADS"] = str(snakemake.threads) + import pandas as pd import sys import math diff --git a/workflow/scripts/hap-diffs.R b/workflow/scripts/hap-diffs.R index c21934805..49236d0b6 100644 --- a/workflow/scripts/hap-diffs.R +++ b/workflow/scripts/hap-diffs.R @@ -4,23 +4,23 @@ library(scales) library(ggforce) library(cowplot) library(dplyr) -#library(splitstackshape) -#library(ggridges) -#library(IRanges) +# library(splitstackshape) +# library(ggridges) +# library(IRanges) library(ggrepel) -#library(ggnewscale) -#library(ggside) +# library(ggnewscale) +# library(ggside) library(glue) -#library("tidylog", warn.conflicts = FALSE) -#library(patchwork) -#library(ggh4x) +# library("tidylog", warn.conflicts = FALSE) +# library(patchwork) +# library(ggh4x) library(tools) -#library(purrr) -#library(reticulate) +# library(purrr) +# library(reticulate) library(ggpubr) -#library(weights) -#library(karyoploteR) -#library(zoo) +# library(weights) +# library(karyoploteR) +# library(zoo) library(scales) library(ggplot2) library(ggforce) @@ -28,61 +28,63 @@ library(grid) options(scipen = 999) -Red="#c1272d" -Indigo="#0000a7" -Yellow="#eecc16" -Teal="#008176" -Gray="#b3b3b3" -FONT_SIZE=8 -MODEL_COLORS = c(PacBio=Indigo, - CNN=Red, - XGB=Yellow, - GMM=Teal, - IPD=Gray, - SEMI="purple", - Revio="#f41c90" # black +Red <- "#c1272d" +Indigo <- "#0000a7" +Yellow <- "#eecc16" +Teal <- "#008176" +Gray <- "#b3b3b3" +FONT_SIZE <- 8 +MODEL_COLORS <- c( + PacBio = Indigo, + CNN = Red, + XGB = Yellow, + GMM = Teal, + IPD = Gray, + SEMI = "purple", + Revio = "#f41c90" # black ) -my_ggsave <- function(file, ...){ - file = glue(file) +my_ggsave <- function(file, ...) { + file <- glue(file) print(file) - ext = file_ext(file) - file_without_ext = tools::file_path_sans_ext(file) - ggsave(glue("tmp.{ext}"), bg='transparent', ...) - cmd = glue("cp tmp.{ext} {file}") - fwrite(last_plot()$data, file=file_without_ext + ".tbl.gz", sep="\t") + ext <- file_ext(file) + file_without_ext <- tools::file_path_sans_ext(file) + ggsave(glue("tmp.{ext}"), bg = "transparent", ...) + cmd <- glue("cp tmp.{ext} {file}") + fwrite(last_plot()$data, file = file_without_ext + ".tbl.gz", sep = "\t") print(cmd) system(cmd) } -my_grid = function(...){ - theme_minimal_grid(font_size=FONT_SIZE, ...) -} +my_grid <- function(...) { + theme_minimal_grid(font_size = FONT_SIZE, ...) +} -my_hgrid = function(...){ - theme_minimal_hgrid(font_size=FONT_SIZE, ...) -} +my_hgrid <- function(...) { + theme_minimal_hgrid(font_size = FONT_SIZE, ...) +} -my_vgrid = function(...){ - theme_minimal_vgrid(font_size=FONT_SIZE, ...) -} +my_vgrid <- function(...) { + theme_minimal_vgrid(font_size = FONT_SIZE, ...) +} reverselog_trans <- function(base = exp(1)) { trans <- function(x) -log(x, base) inv <- function(x) base^(-x) - trans_new(paste0("reverselog-", format(base)), trans, inv, - log_breaks(base = base), - domain = c(1e-100, Inf)) + trans_new(paste0("reverselog-", format(base)), trans, inv, + log_breaks(base = base), + domain = c(1e-100, Inf) + ) } scientific_10 <- function(x) { - is_one = as.numeric(x) == 1 - text = gsub("e", " %*% 10^", scales::scientific_format()(x)) + is_one <- as.numeric(x) == 1 + text <- gsub("e", " %*% 10^", scales::scientific_format()(x)) print(text) - text = str_remove(text, "^1 %\\*% ") # remove leading one + text <- str_remove(text, "^1 %\\*% ") # remove leading one print(text) - text[is_one] = "10^0" - rtn = parse(text=text) + text[is_one] <- "10^0" + rtn <- parse(text = text) rtn } @@ -91,17 +93,17 @@ scientific_10 <- function(x) { # SCRIPT # # -p_threshold=0.05 -in_file=snakemake@input[[1]] -out_file_1=snakemake@output[[1]] -out_file_2=snakemake@output[[2]] -out_file_3=snakemake@output[[3]] -out_file_4=snakemake@output[[4]] +p_threshold <- 0.05 +in_file <- snakemake@input[[1]] +out_file_1 <- snakemake@output[[1]] +out_file_2 <- snakemake@output[[2]] +out_file_3 <- snakemake@output[[3]] +out_file_4 <- snakemake@output[[4]] -df=fread(in_file) %>% +df <- fread(in_file) %>% mutate_at( - c("fire_coverage_H1","fire_coverage_H2","coverage_H1","coverage_H2"), + c("fire_coverage_H1", "fire_coverage_H2", "coverage_H1", "coverage_H2"), as.numeric ) %>% filter(pass_coverage) %>% @@ -112,8 +114,8 @@ df=fread(in_file) %>% !is.na(coverage_H2), ) %>% mutate( - st=peak_start, - en=peak_end, + st = peak_start, + en = peak_end, `#ct` = `#chrom`, ) %>% data.table() @@ -121,30 +123,30 @@ df=fread(in_file) %>% print("Before coverage filter") print(nrow(df)) -# continue -df$hap1_frac_acc = df$fire_coverage_H1/df$coverage_H1 -df$hap2_frac_acc = df$fire_coverage_H2/df$coverage_H2 -df$autosome = "Autosome" -df[`#chrom` == "chrY"]$autosome = "chrY" -df[`#chrom` == "chrX"]$autosome = "chrX" +# continue +df$hap1_frac_acc <- df$fire_coverage_H1 / df$coverage_H1 +df$hap2_frac_acc <- df$fire_coverage_H2 / df$coverage_H2 +df$autosome <- "Autosome" +df[`#chrom` == "chrY"]$autosome <- "chrY" +df[`#chrom` == "chrX"]$autosome <- "chrX" # filter by coverage -sd = 3 -cov = median(df$coverage) -my_min_cov = max(cov*0.5 - sd * sqrt(cov*0.5), 10) -my_max_cov = cov*0.5 + sd * sqrt(cov*0.5) -#my_min_cov = 10 -#my_max_cov = median(df$coverage) +sd <- 3 +cov <- median(df$coverage) +my_min_cov <- max(cov * 0.5 - sd * sqrt(cov * 0.5), 10) +my_max_cov <- cov * 0.5 + sd * sqrt(cov * 0.5) +# my_min_cov = 10 +# my_max_cov = median(df$coverage) -#print(glue("min_cov={my_min_cov} max_cov={my_max_cov} cov={cov}")) +# print(glue("min_cov={my_min_cov} max_cov={my_max_cov} cov={cov}")) -pdf = df %>% +pdf <- df %>% filter(coverage_H1 > 0 & coverage_H2 > 0) %>% mutate( min_cov = my_min_cov, max_cov = my_max_cov, ) %>% - filter(autosome != "chrY" ) %>% + filter(autosome != "chrY") %>% filter(coverage_H1 > min_cov & coverage_H2 > min_cov) %>% filter(coverage_H1 < max_cov & coverage_H2 < max_cov) %>% mutate( @@ -157,7 +159,7 @@ pdf = df %>% print("After coverage filter") print(nrow(pdf)) -if(nrow(pdf)== 0){ +if (nrow(pdf) == 0) { system(glue("touch {out_file_1}")) system(glue("touch {out_file_2}")) system(glue("touch {out_file_3}")) @@ -165,21 +167,21 @@ if(nrow(pdf)== 0){ quit() } -pdf = pdf %>% +pdf <- pdf %>% filter( - hap1_acc >= 0, - hap1_nacc >= 0, - hap2_acc >= 0, - hap2_nacc >= 0, + hap1_acc >= 0, + hap1_nacc >= 0, + hap2_acc >= 0, + hap2_nacc >= 0, ) %>% rowwise() %>% mutate( - p_value=fisher.test(matrix(c(hap1_acc, hap1_nacc, hap2_acc, hap2_nacc),nrow=2))$p.value + p_value = fisher.test(matrix(c(hap1_acc, hap1_nacc, hap2_acc, hap2_nacc), nrow = 2))$p.value ) %>% ungroup() %>% group_by(autosome) %>% mutate( - p_adjust = p.adjust(p_value, method="BH"), + p_adjust = p.adjust(p_value, method = "BH"), ) %>% select(!starts_with("V")) %>% mutate( @@ -194,66 +196,67 @@ print("After p-value adjustment") print(nrow(pdf %>% filter(p_adjust <= p_threshold))) # make the plots -tdf = pdf +tdf <- pdf tdf %>% - ggplot(aes(x=hap1_frac_acc, y=hap2_frac_acc)) + - stat_cor(size=2) + - geom_hex(bins=75) + - geom_abline(aes(intercept=0, slope=1), linetype="dashed")+ - scale_fill_distiller("", palette = "Spectral", trans="log10") + - scale_x_continuous("Paternal accessibility", labels=percent) + - scale_y_continuous("Maternal accessibility", labels=percent) + - #annotation_logticks(sides="lb") + - facet_wrap(~autosome, ncol=2)+ + ggplot(aes(x = hap1_frac_acc, y = hap2_frac_acc)) + + stat_cor(size = 2) + + geom_hex(bins = 75) + + geom_abline(aes(intercept = 0, slope = 1), linetype = "dashed") + + scale_fill_distiller("", palette = "Spectral", trans = "log10") + + scale_x_continuous("H1 accessibility", labels = percent) + + scale_y_continuous("H2 accessibility", labels = percent) + + # annotation_logticks(sides="lb") + + facet_wrap(~autosome, ncol = 2) + my_grid() -my_ggsave(out_file_1, height=3, width=6) +my_ggsave(out_file_1, height = 3, width = 6) -cor_p_threshold = max(tdf[p_adjust <= p_threshold & !is.na(p_value) & !is.na(p_adjust) ]$p_value) +cor_p_threshold <- max(tdf[p_adjust <= p_threshold & !is.na(p_value) & !is.na(p_adjust)]$p_value) print(cor_p_threshold) -y_lim = ceiling(max(max(-log10(tdf$p_value)), max(-log10(tdf$p_adjust)))) -y_by = 1 -if(y_lim > 10){ - y_by = 2 +y_lim <- ceiling(max(max(-log10(tdf$p_value)), max(-log10(tdf$p_adjust)))) +y_by <- 1 +if (y_lim > 10) { + y_by <- 2 } # add p-value col, volcano plot -n=comma(nrow(tdf)) -p = tdf %>% - ggplot(aes(x=diff, y=p_value)) + - geom_hex(bins=100) + scale_fill_distiller("", palette = "Spectral", trans="log10") + - geom_hline(aes(yintercept=(p_threshold)), linetype="dashed", color="darkblue")+ - geom_hline(aes(yintercept=(cor_p_threshold)), linetype="dashed", color="darkred")+ - facet_wrap(~autosome, ncol=2)+ - scale_x_continuous("Difference between paternal and maternal accessibility", labels=percent) + +n <- comma(nrow(tdf)) +p <- tdf %>% + ggplot(aes(x = diff, y = p_value)) + + geom_hex(bins = 100) + + scale_fill_distiller("", palette = "Spectral", trans = "log10") + + geom_hline(aes(yintercept = (p_threshold)), linetype = "dashed", color = "darkblue") + + geom_hline(aes(yintercept = (cor_p_threshold)), linetype = "dashed", color = "darkred") + + facet_wrap(~autosome, ncol = 2) + + scale_x_continuous("Difference between H1 and H2 accessibility", labels = percent) + scale_y_continuous( - glue("p-value (n = {n})"), - #labels=comma, - breaks=10**(-seq(0, y_lim, y_by)), - minor_breaks=10**(-seq(0, y_lim, 0.1)), - trans=reverselog_trans(10), - labels=scientific_10, - ) + + glue("p-value (n = {n})"), + # labels=comma, + breaks = 10**(-seq(0, y_lim, y_by)), + minor_breaks = 10**(-seq(0, y_lim, 0.1)), + trans = reverselog_trans(10), + labels = scientific_10, + ) + my_grid() -my_ggsave(out_file_2, height=3, width=5) +my_ggsave(out_file_2, height = 3, width = 5) -# save the table -fwrite(tdf[tdf$p_value <= 1], out_file_3, sep="\t") +# save the table +fwrite(tdf[tdf$p_value <= 1], out_file_3, sep = "\t") # save the bed9 -# +# tdf %>% mutate( - score = pmin(round(1/p_value), 1000), + score = pmin(round(1 / p_value), 1000), name = paste0( - round(p_value,5), "_", - round(100*hap1_frac_acc,2), "_", - round(100*hap2_frac_acc,2) + round(p_value, 5), "_", + round(100 * hap1_frac_acc, 2), "_", + round(100 * hap2_frac_acc, 2) ), - tst=st+1, - ten=case_when( - p_value<=p_threshold ~ en - 1, - TRUE ~ st+2 + tst = st + 1, + ten = case_when( + p_value <= p_threshold ~ en - 1, + TRUE ~ st + 2 ), - strand=".", + strand = ".", color = case_when( hap1_frac_acc > hap2_frac_acc & p_value > p_threshold ~ "0,0,100", hap1_frac_acc > hap2_frac_acc ~ "0,0,255", @@ -263,4 +266,4 @@ tdf %>% ) ) %>% select(c("#chrom", "st", "en", "name", "score", "strand", "tst", "ten", "color")) %>% - fwrite(out_file_4, sep="\t", quote=F, na=".", row.names=F) + fwrite(out_file_4, sep = "\t", quote = F, na = ".", row.names = F) diff --git a/workflow/scripts/percent-in-clusters.py b/workflow/scripts/percent-in-clusters.py index 4a86c3595..e0ae7437b 100644 --- a/workflow/scripts/percent-in-clusters.py +++ b/workflow/scripts/percent-in-clusters.py @@ -16,7 +16,7 @@ def my_groupby(gdf): real_bp = (df[df.case == "Real"]["length"] * df[df.case == "Real"]["cov"]).sum() df = df.groupby("cov").apply(my_groupby).reset_index() over_expected = df[0][df[0] > 0].sum() -print(f"{over_expected/real_bp:%}\n") +print(f"{over_expected / real_bp:%}\n") # chr1 0 1 12 1 1 1 cov = sys.argv[2] @@ -28,7 +28,7 @@ def my_groupby(gdf): rtn = f"""percent-of-MSPs-preferentially-clustered-along-the-genome\tmin_fdr -{over_expected/real_bp:%}\t{min_fdr} +{over_expected / real_bp:%}\t{min_fdr} """ out = sys.argv[3] open(out, "w").write(rtn)