reverse_locations_if_needed: Reverse modification locations if needed...

In ggDNAvis: 'ggplot2'-Based Tools for Visualising DNA Sequences and Modifications

Reverse modification locations if needed (merge_methylation_with_metadata() helper)

Description

This function takes a vector of condensed modification locations/indices (e.g. c("3,6,9,12", "1,4,7,10")), a vector of directions (which must all be either "forward" or "reverse", not case-sensitive), and a vector of sequence lengths (integers).

Returns a vector of condensed locations where reads that were originally forward are unchanged, and reads that were originally reverse are flipped to now be forward.

Optionally, a numerical offset can be set. If this is left at 0 (the default value), then a CpG assessed for methylation would be reverse-complemented to a CG with the modification information ascribed to the G (as the G is at the location where the actual modified C was on the other strand). However, setting the offset to 1 would shift all of the modification indices by 1 such that the modification is now ascribed to the C of the reverse-strand CG. This is beneficial for visualising the modifications as it ensures consistency between originally-forward and originally-reverse strands by making the modification score associated with each CpG site always be located at the C, but may be misleading for quantitative analysis. Setting the offset to anything other than 0 or 1 should work but may be biologically misleading, so produces a warning.

Called by merge_methylation_with_metadata() to create a forward dataset, alongside reverse_sequence_if_needed(), reverse_quality_if_needed(), and reverse_probabilities_if_needed().

Example:

Forward sequence, with indices of Cs in CpGs numbered:

C C C A G G C G G C G G C G A C C G A
            7     10    13      17

(length = 19, locations = "7,10,13,17", CpGs = 7-8, 10-11, 13-14, 17-18)

Reverse sequence, with indices of C in CpGs numbered:

T C G G T C G C C G C C G C C T G G G
  2       6     9     12

(length = 19, locations = "2,6,9,12", CpGs = 2-3, 6-7, 9-10, 12-13)

As CG reverse-complements to itself, each CpG site has a 1:1 correspondence with a CpG site in the reverse strand. Many methylation calling models assess C-methylation at the C of each CpG. To map the locations from C to C, we take ⁠19 - <index>⁠ such that "7,10,13,17" becomes "12,9,6,2" and "2,6,9,12" becomes "17,13,10,7". The symmetry of CpGs means mapping from C to C is also symmetric. This is achieved by setting offset = 1, as mapping C to C involves shifting position by 1.

Conversely, to map the locations from C to G (i.e. preserving the actual location of each modification, which is required if assessed locations are non-symmetric/don't reverse-complement to themselves like CpGs do), we take ⁠20 - <index>⁠ such that "7,10,13,17" becomes "13,10,7,3" i.e. the indices of the Gs in CpGs in the reverse sequence. Likewise "2,6,9,12" becomes "18,14,11,8" i.e. the indices of the Gs in CpGs in the forward sequence. This is achieved by setting offset = 0, as mapping C to G preserves the actual original position at which each modification was assessed, but changes the base to its complement.

In general, new locations are calculated as ⁠(<length> + 1 - <offset>) - <index>⁠. Of course, output locations are reversed before returning so that they all return in ascending order, as is standard for all location vectors/strings.

If wanting to write reversed sequences to FASTQ via write_modified_fastq(), locations must be symmetric (e.g. CpG) and offset must be set to 1. Asymmetric locations are impossible to write to modified FASTQ once reversed because then e.g. cytosine methylation will be assessed at guanines, which SAMtools can't account for. Symmetrically reversing CpGs via offset = 1 is the only way to fix this.

Usage

reverse_locations_if_needed(
  locations_vector,
  direction_vector,
  length_vector,
  offset = 0
)

Arguments

locations_vector	⁠character vector⁠. The locations to be reversed for each sequence/read. Each read should have one character value, representing a comma-separated list of indices at which modification was assessed along the read e.g. "3,6,9,12" for all the Cs in GGCGGCGGCGGC. These comma-separated characters/strings can be produced from numeric vectors via vector_to_string() and converted back to vectors via string_to_vector().
direction_vector	⁠character vector⁠. Whether each sequence is forward or reverse. Must contain only "forward" and "reverse", but is not case sensitive. Must be the same length as locations_vector and length_vector.
length_vector	⁠integer vector⁠. The length of each sequence. Needed for reversing locations as locations are stored relative to the start of the read i.e. relative to the end of the reverse read. Must be the same length as locations_vector and direction_vector.
offset	integer. How much locations should be shifted by. Defaults to 0. This is important because if a CpG is assessed for methylation at the C, then reverse complementing it will give a methylation score at the G on the reverse-complemented strand. This is the most biologically accurate, but for visualising methylation it may be desired to shift the locations by 1 i.e. to correspond with the C in the reverse-complemented CpG rather than the G, which allows for consistent visualisation between forward and reverse strands. Setting (integer) values other than 0 or 1 will work, but may be biologically misleading so it is not recommended.

Value

⁠character vector⁠. A vector of all forward versions of the input locations vector.

Examples

reverse_locations_if_needed(
    locations_vector = c("7,10,13,17", "2,6,9,12"),
    direction_vector = c("forward", "reverse"),
    length_vector = c(19, 19),
    offset = 0
)

reverse_locations_if_needed(
    locations_vector = c("7,10,13,17", "2,6,9,12"),
    direction_vector = c("forward", "reverse"),
    length_vector = c(19, 19),
    offset = 1
)

ggDNAvis documentation built on Nov. 5, 2025, 7:36 p.m.