Calculating Intron relative distribution
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7.2 years ago

Hi,

I would like to be able to calculate the relative distribution of introns in genes.

A paper from Jeffares et al, 2006 "The biology of intron gain and loss" shows graph of the proportion of introns and their distribution to show the 5' bias in Eukaryote.

It is explained that : relative position of the intron = bp upstream of the intron in coding sequence / total number of bp in coding sequence. Then, they pool it into ten classes.

 

I would like to do something similar for at least the human genome. I am interested in doing this for each chromosome to be able to compare the distribution.

I found that the UCSC has a pretty good database with exon position and I was wondering if it was possible to use this data. I know it would probably require to write a program but I am not a programmer  and the only code I know is R.

Any help is appreciated,

Thank you

 

 

intron UCSC Distribution density • 2.1k views
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Entering edit mode
7.2 years ago
Martin Morgan ★ 1.6k

For a model organism, you could load the TxDb annotation package (see, e.g., TxDb.Hsapiens.UCSC.hg19.knownGene), extract the introns and transcripts from the data, and then calculate the relative offset (of the intron start from the transcript start on the plus strand, for instance)

library(TxDb.Hsapiens.UCSC.hg19.knownGene)
introns <- unlist(intronsByTranscript(
    TxDb.Hsapiens.UCSC.hg19.knownGene, use.names=TRUE))
tx <- transcripts(TxDb.Hsapiens.UCSC.hg19.knownGene,
                  columns=c("tx_name", "TXCHROM"))

idx <- match(names(introns), tx$tx_name)
offset <- ifelse(strand(introns) == "+",
                 start(introns) - start(tx)[idx],
                 end(tx)[idx] - end(introns)) / width(tx)[idx]

The commands are from the GenomicFeatures and GenomicRanges packages. Plot the result

hist(offset)

For less model organisms, a 'TxDb' database can be created with the vignette on the GenomicFeatures landing page.

You could coordinate the offset information with chromosome (with a little bit of trickery to deal with the appropriate factor 'levels' and to focus only on the autosome and sex chromosomes) with

df <- data.frame(offset=offset, chrom=tx$TXCHROM[idx])
lvls <- paste0("chr", c(1:22, "X", "Y"))
df <- df[df$chrom %in% lvls,]

After subsetting factors, it is convenient to drop the unused levels

df$chrom <- factor(df$chrom, lvls)

Visualize the information with, e.g.,

library(lattice)
densityplot(~ offset | chrom, df, plot.points=FALSE)
densityplot(~ offset, group=chrom, df,   
    plot.points=FALSE)

To aggregate several levels, one possibility is along the lines of

levels(df$chrom) <- c(rep("auto", 22), rep("sex", 2))
densityplot(~offset, group=chrom, df, plot.points=FALSE)
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Wow it works pretty well!

I still don't get everything, especially how works the database but I'm looking into it ! Thanks !

However I still can't figure out how to regroup all the autosomes together. I understand I have to deal with the "lvls" but I stuck.

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I revised the final part of the question to separately drop unused factor levels after subsetting, and then to illustrate one way of changing the labels on the new levels

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Fantastic !

It's exactly what I was looking for ! I'm already starting to look for the other database I can use this with.

Thank you again !

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