re-doing chexmix peaks to annotate to nearest downstream orf
Browse files
README.md
CHANGED
|
@@ -81,8 +81,7 @@ features:
|
|
| 81 |
- rossi_2021_af_combined
|
| 82 |
- rossi_2021_af_replicates_mindel
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| 83 |
- rossi_2021_af_combined_mindel
|
| 84 |
-
-
|
| 85 |
-
- yep_filtered_peaks_mindel
|
| 86 |
fields:
|
| 87 |
- name: target_locus_tag
|
| 88 |
dtype: string
|
|
@@ -102,6 +101,22 @@ features:
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|
| 102 |
dtype: int64
|
| 103 |
description: Promoter region end position (1-based, inclusive)
|
| 104 |
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| 105 |
- applies_to:
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| 106 |
- rossi_2021_af_replicates
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| 107 |
- rossi_2021_af_combined
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|
@@ -156,8 +171,6 @@ features:
|
|
| 156 |
- applies_to:
|
| 157 |
- rossi_2021_af_replicates_mindel
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| 158 |
- rossi_2021_af_combined_mindel
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| 159 |
-
- yep_filtered_peaks_yiming
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| 160 |
-
- yep_filtered_peaks_mindel
|
| 161 |
fields:
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| 162 |
- name: width
|
| 163 |
dtype: int64
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|
@@ -187,20 +200,28 @@ features:
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| 187 |
this is a feature from the Mindel data that I have not documented. It is a TODO.
|
| 188 |
|
| 189 |
- applies_to:
|
| 190 |
-
-
|
| 191 |
-
- yep_filtered_peaks_mindel
|
| 192 |
fields:
|
| 193 |
-
- name:
|
| 194 |
dtype: float64
|
| 195 |
description: >-
|
| 196 |
The filtered bed file representing the high confidence ChExMix peaks from
|
| 197 |
-
yeastepigenome.org score. Where multiple peaks
|
| 198 |
-
|
| 199 |
-
combining replicates, the median score across all replicates is taken.
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| 200 |
-
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| 201 |
-
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| 202 |
-
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| 203 |
-
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| 204 |
|
| 205 |
configs:
|
| 206 |
- config_name: rossi_2021_metadata_replicate
|
|
@@ -231,7 +252,7 @@ configs:
|
|
| 231 |
- config_name: rossi_2021_metadata_sample
|
| 232 |
description: Sample-level metadata for combined ChIP-exo experiments including experimental conditions
|
| 233 |
dataset_type: metadata
|
| 234 |
-
applies_to: ["rossi_2021_af_combined", "rossi_2021_af_combined_mindel", "
|
| 235 |
data_files:
|
| 236 |
- split: train
|
| 237 |
path: rossi_2021_metadata_sample.parquet
|
|
@@ -395,47 +416,17 @@ configs:
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|
| 395 |
description: Unique identifier combining regulator and replicates
|
| 396 |
role: sample_id
|
| 397 |
|
| 398 |
-
- config_name:
|
| 399 |
description: >-
|
| 400 |
-
The filtered high confidence peaks reported at yeastepigenome.org
|
| 401 |
-
|
| 402 |
-
|
| 403 |
-
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| 404 |
-
|
| 405 |
-
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| 406 |
-
dataset_type: annotated_features
|
| 407 |
-
genome_resources:
|
| 408 |
-
region_sets:
|
| 409 |
-
Kang:
|
| 410 |
-
path: https://huggingface.co/datasets/BrentLab/yeast_genome_resources/blob/main/yiming_promoters.bed
|
| 411 |
-
join_column: target_locus_tag
|
| 412 |
-
data_files:
|
| 413 |
-
- split: train
|
| 414 |
-
path: yep_filtered_peaks_yiming.parquet
|
| 415 |
-
dataset_info:
|
| 416 |
-
features:
|
| 417 |
-
- name: sample_id
|
| 418 |
-
dtype: int64
|
| 419 |
-
description: Unique identifier for the biological replicate
|
| 420 |
-
role: sample_id
|
| 421 |
-
|
| 422 |
-
- config_name: yep_filtered_peaks_mindel
|
| 423 |
-
description: >-
|
| 424 |
-
The filtered high confidence peaks reported at yeastepigenome.org intersected
|
| 425 |
-
with the Mindel promoter regions. see `scripts/parse_rossi_filtered_peaks.R`
|
| 426 |
-
for more details. NOTE: be sure to read the definition of the score column,
|
| 427 |
-
which is at the top of this README in the common features section, as the
|
| 428 |
-
score has been combined across peaks intersecting the same promoter in the same
|
| 429 |
-
replicate, and then also across replicates.
|
| 430 |
-
dataset_type: annotated_features
|
| 431 |
-
genome_resources:
|
| 432 |
-
region_sets:
|
| 433 |
-
Mindel:
|
| 434 |
-
path: https://huggingface.co/datasets/BrentLab/yeast_genome_resources/blob/main/mindel_promoters.csv.gz
|
| 435 |
-
join_column: target_locus_tag
|
| 436 |
data_files:
|
| 437 |
- split: train
|
| 438 |
-
path:
|
| 439 |
dataset_info:
|
| 440 |
features:
|
| 441 |
- name: sample_id
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|
|
|
| 81 |
- rossi_2021_af_combined
|
| 82 |
- rossi_2021_af_replicates_mindel
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| 83 |
- rossi_2021_af_combined_mindel
|
| 84 |
+
- yep_filtered_peaks_combined
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|
|
|
| 85 |
fields:
|
| 86 |
- name: target_locus_tag
|
| 87 |
dtype: string
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|
|
| 101 |
dtype: int64
|
| 102 |
description: Promoter region end position (1-based, inclusive)
|
| 103 |
|
| 104 |
+
- applies_to:
|
| 105 |
+
- rossi_2021_af_replicates
|
| 106 |
+
- rossi_2021_af_combined
|
| 107 |
+
- rossi_2021_af_replicates_mindel
|
| 108 |
+
- rossi_2021_af_combined_mindel
|
| 109 |
+
fields:
|
| 110 |
+
- name: seqnames
|
| 111 |
+
dtype: string
|
| 112 |
+
description: Chromosome identifier (e.g., chrI, chrII, chrXVI)
|
| 113 |
+
- name: start
|
| 114 |
+
dtype: int64
|
| 115 |
+
description: Promoter region start position (1-based coordinate)
|
| 116 |
+
- name: end
|
| 117 |
+
dtype: int64
|
| 118 |
+
description: Promoter region end position (1-based, inclusive)
|
| 119 |
+
|
| 120 |
- applies_to:
|
| 121 |
- rossi_2021_af_replicates
|
| 122 |
- rossi_2021_af_combined
|
|
|
|
| 171 |
- applies_to:
|
| 172 |
- rossi_2021_af_replicates_mindel
|
| 173 |
- rossi_2021_af_combined_mindel
|
|
|
|
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|
|
| 174 |
fields:
|
| 175 |
- name: width
|
| 176 |
dtype: int64
|
|
|
|
| 200 |
this is a feature from the Mindel data that I have not documented. It is a TODO.
|
| 201 |
|
| 202 |
- applies_to:
|
| 203 |
+
- yep_filtered_peaks_combined
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|
|
|
| 204 |
fields:
|
| 205 |
+
- name: peak_score
|
| 206 |
dtype: float64
|
| 207 |
description: >-
|
| 208 |
The filtered bed file representing the high confidence ChExMix peaks from
|
| 209 |
+
yeastepigenome.org score. Where multiple peaks are within 500bp upstream of a target
|
| 210 |
+
ORF, we take the median peak score. Replicates are then combined. When
|
| 211 |
+
combining replicates, the median score across all replicates is taken.
|
| 212 |
+
- name: n_peaks
|
| 213 |
+
dtype: float64
|
| 214 |
+
description: >-
|
| 215 |
+
The number of peaks annotated to the same target. This is both within replicates and
|
| 216 |
+
across (ie, there might be 3 peaks in 2 replicates for a total of 6 in this data)
|
| 217 |
+
- name: max_distance
|
| 218 |
+
dtype: float64
|
| 219 |
+
description: >-
|
| 220 |
+
The maximum distance a peak in the set (see n_peaks) is from the ORF
|
| 221 |
+
- name: peak_n_replicates
|
| 222 |
+
dtype: float64
|
| 223 |
+
description: >-
|
| 224 |
+
The number of replicates which have peaks for this target
|
| 225 |
|
| 226 |
configs:
|
| 227 |
- config_name: rossi_2021_metadata_replicate
|
|
|
|
| 252 |
- config_name: rossi_2021_metadata_sample
|
| 253 |
description: Sample-level metadata for combined ChIP-exo experiments including experimental conditions
|
| 254 |
dataset_type: metadata
|
| 255 |
+
applies_to: ["rossi_2021_af_combined", "rossi_2021_af_combined_mindel", "yep_filtered_peaks_combined"]
|
| 256 |
data_files:
|
| 257 |
- split: train
|
| 258 |
path: rossi_2021_metadata_sample.parquet
|
|
|
|
| 416 |
description: Unique identifier combining regulator and replicates
|
| 417 |
role: sample_id
|
| 418 |
|
| 419 |
+
- config_name: yep_filtered_peaks_combined
|
| 420 |
description: >-
|
| 421 |
+
The filtered high confidence peaks reported at yeastepigenome.org. The peaks
|
| 422 |
+
are annotated to the nearest downstream gene (not dubious). Peaks within 500
|
| 423 |
+
bp of the ORF are keep. Within a given replicate, if a target has more than 1
|
| 424 |
+
peak, then the median score is taken. Replicates are combined in the same way,
|
| 425 |
+
with the median target score across replicates. see
|
| 426 |
+
`scripts/parse_rossi_filtered_peaks.R` for more details.
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|
| 427 |
data_files:
|
| 428 |
- split: train
|
| 429 |
+
path: yep_filtered_peaks_combined.parquet
|
| 430 |
dataset_info:
|
| 431 |
features:
|
| 432 |
- name: sample_id
|
scripts/CopyOfparse_rossi_filtered_peaks.R
ADDED
|
@@ -0,0 +1,210 @@
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| 1 |
+
# fetch the zip files from the yeastepigenome.org server
|
| 2 |
+
# https://www.datacommons.psu.edu/download/eberly/pughlab/yeast-epigenome-project/
|
| 3 |
+
# and then extract them
|
| 4 |
+
# with
|
| 5 |
+
#!/usr/bin/env bash
|
| 6 |
+
# fetch_yep_peaks.sh
|
| 7 |
+
#
|
| 8 |
+
# Extracts *_chexmix_filtered_peaks.bed from each *_YEP.zip in ZIP_DIR.
|
| 9 |
+
# Output: yep_filtered_peaks/ in the current working directory.
|
| 10 |
+
#
|
| 11 |
+
# set -euo pipefail
|
| 12 |
+
#
|
| 13 |
+
# ZIP_DIR="${1:-/lts/mblab/downloaded_data/chipexo/yeastepigenome_data}"
|
| 14 |
+
# OUT_DIR="$(pwd)/yep_filtered_peaks"
|
| 15 |
+
#
|
| 16 |
+
# if [[ ! -d "$ZIP_DIR" ]]; then
|
| 17 |
+
# echo "[error] ZIP_DIR not found: $ZIP_DIR" >&2
|
| 18 |
+
# exit 1
|
| 19 |
+
# fi
|
| 20 |
+
#
|
| 21 |
+
# mkdir -p "$OUT_DIR"
|
| 22 |
+
#
|
| 23 |
+
# COUNT=0; SKIP=0; FAIL=0
|
| 24 |
+
#
|
| 25 |
+
# for zip_path in "$ZIP_DIR"/*_YEP.zip; do
|
| 26 |
+
# zip_name=$(basename "$zip_path")
|
| 27 |
+
# sample_id="${zip_name%_YEP.zip}"
|
| 28 |
+
# bed_name="${sample_id}_chexmix_filtered_peaks.bed"
|
| 29 |
+
# dest="$OUT_DIR/$bed_name"
|
| 30 |
+
#
|
| 31 |
+
# if [[ -f "$dest" ]]; then
|
| 32 |
+
# echo "[skip] $bed_name already exists"
|
| 33 |
+
# (( SKIP++ )) || true
|
| 34 |
+
# continue
|
| 35 |
+
# fi
|
| 36 |
+
#
|
| 37 |
+
# # file path inside the zip may vary — find it
|
| 38 |
+
# bed_in_zip=$(unzip -Z1 "$zip_path" 2>/dev/null | grep -m1 "${bed_name}$" || true)
|
| 39 |
+
#
|
| 40 |
+
# if [[ -z "$bed_in_zip" ]]; then
|
| 41 |
+
# echo "[warn] $bed_name not found in $zip_name — skipping" >&2
|
| 42 |
+
# (( FAIL++ )) || true
|
| 43 |
+
# continue
|
| 44 |
+
# fi
|
| 45 |
+
#
|
| 46 |
+
# echo "[ext ] $zip_name → $bed_name"
|
| 47 |
+
# unzip -p "$zip_path" "$bed_in_zip" > "$dest"
|
| 48 |
+
# (( COUNT++ )) || true
|
| 49 |
+
# done
|
| 50 |
+
#
|
| 51 |
+
# echo ""
|
| 52 |
+
# echo "[done] extracted=$COUNT skipped=$SKIP failed=$FAIL"
|
| 53 |
+
# echo "[done] output in: $OUT_DIR"
|
| 54 |
+
|
| 55 |
+
# the following can be run on the output
|
| 56 |
+
|
| 57 |
+
library(tidyverse)
|
| 58 |
+
library(arrow)
|
| 59 |
+
library(GenomicRanges)
|
| 60 |
+
library(here)
|
| 61 |
+
|
| 62 |
+
#' Convert BED format data frame to GRanges
|
| 63 |
+
#'
|
| 64 |
+
#' Handles coordinate system conversion from 0-indexed half-open BED format
|
| 65 |
+
#' to 1-indexed closed GenomicRanges format
|
| 66 |
+
#'
|
| 67 |
+
#' @param bed_df Data frame with chr, start, end columns in BED format (0-indexed, half-open)
|
| 68 |
+
#' @param zero_indexed Logical, whether input is 0-indexed (default: TRUE)
|
| 69 |
+
#' @return GRanges object
|
| 70 |
+
bed_to_granges <- function(bed_df, zero_indexed = TRUE) {
|
| 71 |
+
|
| 72 |
+
if (!all(c("chr", "start", "end") %in% names(bed_df))) {
|
| 73 |
+
stop("bed_df must have columns: chr, start, end")
|
| 74 |
+
}
|
| 75 |
+
|
| 76 |
+
# Convert from 0-indexed half-open [start, end) to 1-indexed closed [start, end]
|
| 77 |
+
if (zero_indexed) {
|
| 78 |
+
gr_start <- bed_df$start + 1
|
| 79 |
+
gr_end <- bed_df$end
|
| 80 |
+
} else {
|
| 81 |
+
gr_start <- bed_df$start
|
| 82 |
+
gr_end <- bed_df$end
|
| 83 |
+
}
|
| 84 |
+
|
| 85 |
+
# Create GRanges object (strand-agnostic for calling cards)
|
| 86 |
+
gr <- GenomicRanges::GRanges(
|
| 87 |
+
seqnames = bed_df$chr,
|
| 88 |
+
ranges = IRanges::IRanges(start = gr_start, end = gr_end),
|
| 89 |
+
strand = "*"
|
| 90 |
+
)
|
| 91 |
+
|
| 92 |
+
# Add any additional metadata columns
|
| 93 |
+
extra_cols <- setdiff(names(bed_df), c("chr", "start", "end", "strand"))
|
| 94 |
+
if (length(extra_cols) > 0) {
|
| 95 |
+
GenomicRanges::mcols(gr) <- bed_df[, extra_cols, drop = FALSE]
|
| 96 |
+
}
|
| 97 |
+
|
| 98 |
+
return(gr)
|
| 99 |
+
}
|
| 100 |
+
|
| 101 |
+
#' Annotate genomic regions with ChExMix peak scores
|
| 102 |
+
#'
|
| 103 |
+
#' Intersects a set of genomic regions (e.g. promoters) with ChExMix filtered
|
| 104 |
+
#' peaks and assigns each region a score. Regions overlapping a single peak
|
| 105 |
+
#' receive that peak's score; regions overlapping multiple peaks receive the
|
| 106 |
+
#' median score across all overlapping peaks. Non-overlapping regions are
|
| 107 |
+
#' scored 0.
|
| 108 |
+
#'
|
| 109 |
+
#' @param peaks_df Data frame with BED-format peak calls (0-indexed, half-open)
|
| 110 |
+
#' and a \code{score} column, as produced by \code{read_in_filtered_bed}.
|
| 111 |
+
#' @param regions_gr GRanges object of genomic regions to annotate
|
| 112 |
+
#' (e.g. an element of \code{regions_list}).
|
| 113 |
+
#'
|
| 114 |
+
#' @return A copy of \code{regions_gr} with an additional metadata column
|
| 115 |
+
#' \code{score}: the peak score for overlapping regions, 0 otherwise.
|
| 116 |
+
#'
|
| 117 |
+
#' @seealso \code{\link{bed_to_granges}}, \code{\link{read_in_filtered_bed}}
|
| 118 |
+
peaks_to_promoter_scores <- function(peaks_df, regions_gr) {
|
| 119 |
+
|
| 120 |
+
peaks_gr <- bed_to_granges(peaks_df)
|
| 121 |
+
|
| 122 |
+
hits <- GenomicRanges::findOverlaps(regions_gr, peaks_gr)
|
| 123 |
+
|
| 124 |
+
# for each hit, pull the peak score
|
| 125 |
+
scores_df <- tibble(
|
| 126 |
+
region_idx = S4Vectors::queryHits(hits),
|
| 127 |
+
score = peaks_df$score[S4Vectors::subjectHits(hits)]
|
| 128 |
+
) |>
|
| 129 |
+
summarise(score = median(score), .by = region_idx)
|
| 130 |
+
|
| 131 |
+
# initialize all promoters to 0, fill in overlapping ones
|
| 132 |
+
result <- rep(0, length(regions_gr))
|
| 133 |
+
result[scores_df$region_idx] <- scores_df$score
|
| 134 |
+
|
| 135 |
+
regions_gr_tmp <- regions_gr
|
| 136 |
+
regions_gr_tmp$score <- result
|
| 137 |
+
regions_gr_tmp
|
| 138 |
+
}
|
| 139 |
+
|
| 140 |
+
|
| 141 |
+
rossi_meta = arrow::read_parquet("~/code/hf/rossi_2021/rossi_2021_metadata.parquet")
|
| 142 |
+
chrmap = read_csv("~/code/hf/yeast_genome_resources/chrmap.csv.gz")
|
| 143 |
+
|
| 144 |
+
brentlab_features = read_csv("~/code/hf/yeast_genome_resources/brentlab_features.csv.gz")
|
| 145 |
+
|
| 146 |
+
# Load regions
|
| 147 |
+
regions_list = list(
|
| 148 |
+
yiming = read_tsv(
|
| 149 |
+
"~/code/hf/yeast_genome_resources/yiming_promoters.bed",
|
| 150 |
+
col_names = c('chr', 'start', 'end', 'locus_tag', 'score', 'strand')) %>%
|
| 151 |
+
bed_to_granges(),
|
| 152 |
+
mindel = arrow::read_parquet("~/code/hf/yeast_genome_resources/mindel_promoters.parquet") |>
|
| 153 |
+
bed_to_granges()
|
| 154 |
+
)
|
| 155 |
+
|
| 156 |
+
filtered_bed = list.files(here("data/yep_filtered_peaks"), full.names = TRUE)
|
| 157 |
+
names(filtered_bed) = str_extract(basename(filtered_bed), "^\\d+")
|
| 158 |
+
|
| 159 |
+
read_in_filtered_bed = function(path){
|
| 160 |
+
read_tsv(path, col_names = c('chr', 'start', 'end', 'source', 'score')) |>
|
| 161 |
+
left_join(dplyr::select(chrmap, chr, ucsc)) |>
|
| 162 |
+
dplyr::select(-chr) |>
|
| 163 |
+
dplyr::rename(chr = ucsc) |>
|
| 164 |
+
dplyr::relocate(chr)
|
| 165 |
+
}
|
| 166 |
+
|
| 167 |
+
filtered_bed_df_list = map(filtered_bed, read_in_filtered_bed)
|
| 168 |
+
|
| 169 |
+
filtered_bed_df_list_no_null = filtered_bed_df_list[unlist(map(filtered_bed_df_list, ~nrow(.) > 0))]
|
| 170 |
+
|
| 171 |
+
filtered_bed_df_list_no_null_in_meta = filtered_bed_df_list_no_null[
|
| 172 |
+
as.integer(names(filtered_bed_df_list_no_null)) %in% rossi_meta$yeastepigenome_id]
|
| 173 |
+
|
| 174 |
+
af_list = list(
|
| 175 |
+
yiming = map(filtered_bed_df_list_no_null_in_meta, peaks_to_promoter_scores, regions_list$yiming),
|
| 176 |
+
mindel = map(filtered_bed_df_list_no_null_in_meta, peaks_to_promoter_scores, regions_list$mindel)
|
| 177 |
+
)
|
| 178 |
+
|
| 179 |
+
af_list_dfs = list(
|
| 180 |
+
yiming = bind_rows(map(af_list$yiming, as_tibble), .id = "yeastepigenome_id") |>
|
| 181 |
+
mutate(yeastepigenome_id = as.integer(yeastepigenome_id)) |>
|
| 182 |
+
left_join(dplyr::select(brentlab_features, locus_tag, symbol)) |>
|
| 183 |
+
dplyr::rename(target_locus_tag = locus_tag, target_symbol = symbol) |>
|
| 184 |
+
left_join(rossi_meta) |>
|
| 185 |
+
group_by(sample_id, seqnames, start, end, width, strand, target_locus_tag, target_symbol) |>
|
| 186 |
+
reframe(score = median(score)) |>
|
| 187 |
+
group_by(sample_id) |>
|
| 188 |
+
arrange(desc(score)),
|
| 189 |
+
mindel = bind_rows(map(af_list$mindel, as_tibble), .id = "yeastepigenome_id") |>
|
| 190 |
+
mutate(yeastepigenome_id = as.integer(yeastepigenome_id)) |>
|
| 191 |
+
left_join(rossi_meta) |>
|
| 192 |
+
group_by(sample_id, seqnames, start, end, width, strand, target_locus_tag, target_symbol) |>
|
| 193 |
+
reframe(score = median(score)) |>
|
| 194 |
+
group_by(sample_id) |>
|
| 195 |
+
arrange(desc(score))
|
| 196 |
+
)
|
| 197 |
+
|
| 198 |
+
# arrow::write_parquet(
|
| 199 |
+
# af_list_dfs$yiming,
|
| 200 |
+
# "~/code/hf/rossi_2021/yep_filtered_peaks_yiming.parquet",
|
| 201 |
+
# compression = "zstd",
|
| 202 |
+
# write_statistics = TRUE
|
| 203 |
+
# )
|
| 204 |
+
#
|
| 205 |
+
# arrow::write_parquet(
|
| 206 |
+
# af_list_dfs$mindel,
|
| 207 |
+
# "~/code/hf/rossi_2021/yep_filtered_peaks_mindel.parquet",
|
| 208 |
+
# compression = "zstd",
|
| 209 |
+
# write_statistics = TRUE
|
| 210 |
+
# )
|
scripts/parse_rossi_filtered_peaks.R
CHANGED
|
@@ -98,60 +98,96 @@ bed_to_granges <- function(bed_df, zero_indexed = TRUE) {
|
|
| 98 |
return(gr)
|
| 99 |
}
|
| 100 |
|
| 101 |
-
#'
|
| 102 |
#'
|
| 103 |
-
#'
|
| 104 |
-
#'
|
| 105 |
-
#'
|
| 106 |
-
#'
|
| 107 |
-
#'
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
| 108 |
#'
|
| 109 |
#' @param peaks_df Data frame with BED-format peak calls (0-indexed, half-open)
|
| 110 |
#' and a \code{score} column, as produced by \code{read_in_filtered_bed}.
|
| 111 |
-
#' @param
|
| 112 |
-
#'
|
|
|
|
|
|
|
| 113 |
#'
|
| 114 |
-
#' @return A
|
| 115 |
-
#' \code{
|
|
|
|
|
|
|
| 116 |
#'
|
| 117 |
-
#' @seealso \code{\link{bed_to_granges}}, \code{\link{
|
| 118 |
-
|
| 119 |
|
| 120 |
peaks_gr <- bed_to_granges(peaks_df)
|
| 121 |
|
| 122 |
-
|
| 123 |
-
|
| 124 |
-
|
| 125 |
-
|
| 126 |
-
|
| 127 |
-
|
| 128 |
-
|
| 129 |
-
|
| 130 |
-
|
| 131 |
-
|
| 132 |
-
|
| 133 |
-
|
| 134 |
-
|
| 135 |
-
|
| 136 |
-
|
| 137 |
-
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
| 138 |
}
|
| 139 |
|
| 140 |
|
| 141 |
rossi_meta = arrow::read_parquet("~/code/hf/rossi_2021/rossi_2021_metadata.parquet")
|
| 142 |
chrmap = read_csv("~/code/hf/yeast_genome_resources/chrmap.csv.gz")
|
| 143 |
|
| 144 |
-
brentlab_features =
|
| 145 |
|
| 146 |
-
#
|
| 147 |
-
|
| 148 |
-
yiming = read_tsv(
|
| 149 |
-
"~/code/hf/yeast_genome_resources/yiming_promoters.bed",
|
| 150 |
-
col_names = c('chr', 'start', 'end', 'locus_tag', 'score', 'strand')) %>%
|
| 151 |
-
bed_to_granges(),
|
| 152 |
-
mindel = arrow::read_parquet("~/code/hf/yeast_genome_resources/mindel_promoters.parquet") |>
|
| 153 |
-
bed_to_granges()
|
| 154 |
-
)
|
| 155 |
|
| 156 |
filtered_bed = list.files(here("data/yep_filtered_peaks"), full.names = TRUE)
|
| 157 |
names(filtered_bed) = str_extract(basename(filtered_bed), "^\\d+")
|
|
@@ -171,40 +207,24 @@ filtered_bed_df_list_no_null = filtered_bed_df_list[unlist(map(filtered_bed_df_l
|
|
| 171 |
filtered_bed_df_list_no_null_in_meta = filtered_bed_df_list_no_null[
|
| 172 |
as.integer(names(filtered_bed_df_list_no_null)) %in% rossi_meta$yeastepigenome_id]
|
| 173 |
|
| 174 |
-
|
| 175 |
-
|
| 176 |
-
|
| 177 |
-
|
| 178 |
-
|
| 179 |
-
|
| 180 |
-
|
| 181 |
-
|
| 182 |
-
|
| 183 |
-
|
| 184 |
-
|
| 185 |
-
|
| 186 |
-
|
| 187 |
-
|
| 188 |
-
arrange(desc(score)),
|
| 189 |
-
mindel = bind_rows(map(af_list$mindel, as_tibble), .id = "yeastepigenome_id") |>
|
| 190 |
-
mutate(yeastepigenome_id = as.integer(yeastepigenome_id)) |>
|
| 191 |
-
left_join(rossi_meta) |>
|
| 192 |
-
group_by(sample_id, seqnames, start, end, width, strand, target_locus_tag, target_symbol) |>
|
| 193 |
-
reframe(score = median(score)) |>
|
| 194 |
-
group_by(sample_id) |>
|
| 195 |
-
arrange(desc(score))
|
| 196 |
-
)
|
| 197 |
|
| 198 |
# arrow::write_parquet(
|
| 199 |
-
#
|
| 200 |
-
# "~/code/hf/rossi_2021/
|
| 201 |
-
# compression = "zstd",
|
| 202 |
-
# write_statistics = TRUE
|
| 203 |
-
# )
|
| 204 |
-
#
|
| 205 |
-
# arrow::write_parquet(
|
| 206 |
-
# af_list_dfs$mindel,
|
| 207 |
-
# "~/code/hf/rossi_2021/yep_filtered_peaks_mindel.parquet",
|
| 208 |
# compression = "zstd",
|
| 209 |
# write_statistics = TRUE
|
| 210 |
# )
|
|
|
|
| 98 |
return(gr)
|
| 99 |
}
|
| 100 |
|
| 101 |
+
#' Build a GRanges of protein-coding gene features for peak annotation
|
| 102 |
#'
|
| 103 |
+
#' Restricts the brentlab feature set to protein-coding genes, excluding
|
| 104 |
+
#' dubious ORFs, and returns a strand-aware GRanges carrying the locus tag
|
| 105 |
+
#' and gene symbol as metadata. Coordinates in the feature table are 1-indexed
|
| 106 |
+
#' closed (SGD convention) and are used directly. Also removes mito genes
|
| 107 |
+
#'
|
| 108 |
+
#' @param features_df Data frame from \code{brentlab_features.parquet} with
|
| 109 |
+
#' columns chr, start, end, strand, type, locus_tag, symbol, note.
|
| 110 |
+
#' @return GRanges of retained gene features with metadata columns
|
| 111 |
+
#' \code{locus_tag} and \code{symbol}.
|
| 112 |
+
prepare_gene_features <- function(features_df) {
|
| 113 |
+
|
| 114 |
+
genes <- features_df |>
|
| 115 |
+
dplyr::filter(type == "gene",
|
| 116 |
+
stringr::str_detect(note, stringr::fixed("dubious", ignore_case = TRUE),
|
| 117 |
+
negate = TRUE),
|
| 118 |
+
chr != "chrM")
|
| 119 |
+
|
| 120 |
+
GenomicRanges::GRanges(
|
| 121 |
+
seqnames = genes$chr,
|
| 122 |
+
ranges = IRanges::IRanges(start = genes$start, end = genes$end),
|
| 123 |
+
strand = genes$strand,
|
| 124 |
+
locus_tag = genes$locus_tag,
|
| 125 |
+
symbol = genes$symbol
|
| 126 |
+
)
|
| 127 |
+
}
|
| 128 |
+
|
| 129 |
+
#' Annotate ChExMix peaks to the nearest downstream gene feature
|
| 130 |
+
#'
|
| 131 |
+
#' Assigns each peak to its nearest downstream feature by genomic coordinate
|
| 132 |
+
#' (strand-agnostic, since peaks carry no strand) and records the gap distance.
|
| 133 |
+
#' This proceeds in two rounds:
|
| 134 |
+
#' \enumerate{
|
| 135 |
+
#' \item Every peak is annotated to the nearest downstream gene and the
|
| 136 |
+
#' distance to that gene is computed. Peaks whose nearest downstream gene
|
| 137 |
+
#' lies farther than \code{nbp} (or have no downstream gene) are dropped.
|
| 138 |
+
#' \item Retained peak-to-gene assignments are aggregated by target gene,
|
| 139 |
+
#' and the median peak score across the set of peaks annotated to a gene is
|
| 140 |
+
#' reported as that gene's \code{peak_score}.
|
| 141 |
+
#' }
|
| 142 |
#'
|
| 143 |
#' @param peaks_df Data frame with BED-format peak calls (0-indexed, half-open)
|
| 144 |
#' and a \code{score} column, as produced by \code{read_in_filtered_bed}.
|
| 145 |
+
#' @param features_gr GRanges of gene features, as produced by
|
| 146 |
+
#' \code{prepare_gene_features}.
|
| 147 |
+
#' @param nbp Maximum allowed distance (bp) between a peak and its nearest
|
| 148 |
+
#' downstream gene for the peak to be retained. Default 500.
|
| 149 |
#'
|
| 150 |
+
#' @return A tibble with one row per annotated gene and columns
|
| 151 |
+
#' \code{target_locus_tag}, \code{target_symbol}, \code{peak_score} (median
|
| 152 |
+
#' peak score across annotated peaks) and \code{n_peaks} (number of peaks
|
| 153 |
+
#' annotated to the gene).
|
| 154 |
#'
|
| 155 |
+
#' @seealso \code{\link{bed_to_granges}}, \code{\link{prepare_gene_features}}
|
| 156 |
+
annotate_peaks_to_features <- function(peaks_df, features_gr, nbp = 500) {
|
| 157 |
|
| 158 |
peaks_gr <- bed_to_granges(peaks_df)
|
| 159 |
|
| 160 |
+
# round 1: nearest downstream feature (strand-agnostic) and its distance
|
| 161 |
+
ds_idx <- GenomicRanges::precede(peaks_gr, features_gr, ignore.strand = TRUE)
|
| 162 |
+
keep <- !is.na(ds_idx)
|
| 163 |
+
|
| 164 |
+
peaks_gr <- peaks_gr[keep]
|
| 165 |
+
feat_hits <- features_gr[ds_idx[keep]]
|
| 166 |
+
|
| 167 |
+
round1 <- tibble::tibble(
|
| 168 |
+
score = peaks_df$score[keep],
|
| 169 |
+
target_locus_tag = feat_hits$locus_tag,
|
| 170 |
+
target_symbol = feat_hits$symbol,
|
| 171 |
+
distance = GenomicRanges::distance(peaks_gr, feat_hits,
|
| 172 |
+
ignore.strand = TRUE)) |>
|
| 173 |
+
dplyr::filter(distance <= nbp)
|
| 174 |
+
|
| 175 |
+
# round 2: aggregate peaks per target gene, median score as the peak score
|
| 176 |
+
round1 |>
|
| 177 |
+
dplyr::group_by(target_locus_tag, target_symbol) |>
|
| 178 |
+
dplyr::reframe(peak_score = median(score),
|
| 179 |
+
n_peaks = dplyr::n(),
|
| 180 |
+
max_distance = max(distance))
|
| 181 |
}
|
| 182 |
|
| 183 |
|
| 184 |
rossi_meta = arrow::read_parquet("~/code/hf/rossi_2021/rossi_2021_metadata.parquet")
|
| 185 |
chrmap = read_csv("~/code/hf/yeast_genome_resources/chrmap.csv.gz")
|
| 186 |
|
| 187 |
+
brentlab_features = arrow::read_parquet("~/code/hf/yeast_genome_resources/brentlab_features.parquet")
|
| 188 |
|
| 189 |
+
# protein-coding gene features used to annotate peaks (excludes dubious ORFs)
|
| 190 |
+
gene_features_gr = prepare_gene_features(brentlab_features)
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
| 191 |
|
| 192 |
filtered_bed = list.files(here("data/yep_filtered_peaks"), full.names = TRUE)
|
| 193 |
names(filtered_bed) = str_extract(basename(filtered_bed), "^\\d+")
|
|
|
|
| 207 |
filtered_bed_df_list_no_null_in_meta = filtered_bed_df_list_no_null[
|
| 208 |
as.integer(names(filtered_bed_df_list_no_null)) %in% rossi_meta$yeastepigenome_id]
|
| 209 |
|
| 210 |
+
# annotate each sample's peaks to nearest downstream genes within 500bp
|
| 211 |
+
annotated = map(filtered_bed_df_list_no_null_in_meta,
|
| 212 |
+
annotate_peaks_to_features, gene_features_gr, nbp = 500)
|
| 213 |
+
|
| 214 |
+
annotated_df = bind_rows(annotated, .id = "yeastepigenome_id") |>
|
| 215 |
+
mutate(yeastepigenome_id = as.integer(yeastepigenome_id)) |>
|
| 216 |
+
left_join(rossi_meta, by = "yeastepigenome_id") |>
|
| 217 |
+
group_by(sample_id, target_locus_tag, target_symbol) |>
|
| 218 |
+
reframe(
|
| 219 |
+
peak_score = median(peak_score),
|
| 220 |
+
n_peaks = sum(n_peaks),
|
| 221 |
+
max_distance = max(max_distance),
|
| 222 |
+
peak_n_replicates = n()) |>
|
| 223 |
+
arrange(desc(peak_score), .by_group = TRUE)
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
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|
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|
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|
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|
|
| 224 |
|
| 225 |
# arrow::write_parquet(
|
| 226 |
+
# annotated_df,
|
| 227 |
+
# "~/code/hf/rossi_2021/yep_filtered_peaks_combined.parquet",
|
|
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|
|
|
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|
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|
|
| 228 |
# compression = "zstd",
|
| 229 |
# write_statistics = TRUE
|
| 230 |
# )
|
yep_filtered_peaks_mindel.parquet → yep_filtered_peaks_combined.parquet
RENAMED
|
@@ -1,3 +1,3 @@
|
|
| 1 |
version https://git-lfs.github.com/spec/v1
|
| 2 |
-
oid sha256:
|
| 3 |
-
size
|
|
|
|
| 1 |
version https://git-lfs.github.com/spec/v1
|
| 2 |
+
oid sha256:201410f57fe0368cf14ab1dce12192792293f885517d31f86a07494c9ef86d2a
|
| 3 |
+
size 527546
|
yep_filtered_peaks_yiming.parquet
DELETED
|
@@ -1,3 +0,0 @@
|
|
| 1 |
-
version https://git-lfs.github.com/spec/v1
|
| 2 |
-
oid sha256:cbd1506072d97198bce01b497cb93a33a79772843bd6fa22bfe5dd04e3037795
|
| 3 |
-
size 30745732
|
|
|
|
|
|
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|
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|
|