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add info about scan for start
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elileka authored May 1, 2024
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25 changes: 22 additions & 3 deletions README.md
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Expand Up @@ -103,7 +103,7 @@ This module will extract all putative protein fragments from each contig and str
Since this step involves a search, it is the most time-demanding of all analyses steps. Upon completion, it will output a database (contigs are keys), where each line contains information about a **TCS** and its exon (multi-exon **TCS**s will span several lines).


#### Optional calling of sub-optimal exon sets:
#### OPTIONAL - calling of sub-optimal exon sets:

By default, MetaEuk calls a single and optimal compatible exon set from each **C** & **S** for each **T**. If you are interested in calling several matches to a certain **T** from each **C** & **S** (for example, to look for **gene duplications**), you can change the default value of ```max-exon-sets``` to the number of sets to look for (from version 6). A few important notes:

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#### The MetaEuk header:

The header is composed of several sections, separated by pipes ('|'):
The basic header is composed of several sections, separated by pipes ('|'):

*>T_acc|C_acc|S|bitscore|E-Value|number_exons|low_coord|high_coord|exon1_coords|exon2_coords|...*

Expand All @@ -148,12 +148,31 @@ Example header (two exons on the minus strand):

*>protein_acc|contig_acc|-|1146|0|2|3|1875|1875[1875]:970[970]:906[906]|893[869]:3[3]:891[867]*

Optionally, by setting the flag `--write-frag-coords 1`, information about the position of stop codons will be added to the output. In this case the exon_coords will be given in the following structure:
##### OPTIONAL - adding information about stop codon positions:
By setting the flag `--write-frag-coords 1`, information about the position of stop codons will be added to the output. In this case the exon_coords will be given in the following structure:

*[fragment_low]low[taken_low]:[fragment_high]high[taken_high]:nucleotide_length[taken_nucleotide_length]*

In its initial stage, MetaEuk extracts putative coding fragments between stop codons. It later discovers exons within them by matching targets. The fragment coordinates in square brackets refer to the original fragment in which the exon was found. In addition to reporting these coordinates, MetaEuk will print the stop codon (`*` in the protein output) right at the end of the last exon, if it exists.

##### OPTIONAL - scanning for start codon before the first exon:
By default (`--len-scan-for-start 0`), MetaEuk only reports parts of the contig that match a target. In case of fragmented targets or very distantly-related targets, it can therefore produce predictions, which do not start with a methionine. By setting `--len-scan-for-start` to a positive number, e.g., 50, MetaEuk will scan up-to 50 nucleotides (16 codons) before the first exon of each prediction (upstream for predictions on the plus strand, downstream - for minus).

The scan will be in the same frame as the first exon and not beyond its stop codon border. Within this "legal" window, the scan will finish at the closest methionine to the first exon's matched start. The fragment from the found ATG until the first exon's matched start will be padded to the reported sequence. In the case of predictions on the plus strand, the *low_coord* value (7th field) will be updated to a lower value and the length of the padded fragment will be reported in square brackets. If the scan is turned on but no padding occurred (if the prediction already started with methionine or if no ATG was found), then the *low_coord* value will remain the same, followed by 0 in square brackets. For predictions on the minus strand, the change will be to *high_coord* (8th field). All other fields, including the exon fields, will remain unchanged. Examples:

*>protein_acc|contig_acc|+|784|1.213e-233|4|100[18]|1444|...*
Here, six codons including ATG, were padded before the first exon of a prediction on the plus strand, which starts at position 118 (100+18).

*>protein_acc|contig_acc|-|499|7.54e-148|2|100|911[12]|...*
Here, four codons including ATG, were padded before the first exon of a prediction on the minus strand, which starts at position 899 (911-12).

*>protein_acc|contig_acc|-|499|7.54e-148|2|100|899[0]|...*
Here, no padding occurred, but the scan option was set to a positive number.


Of note, for simplicity, MetaEuk considers only ATG as a start for this scan.


##### The MetaEuk GFF:

In addition to writing a Fasta file, MetaEuk writes a GFF file. Please note that GFF is not perfectly suitable for MetaEuk because MetaEuk doesn't predict non-coding regions. This means that the MetaEuk gene starts and ends where the first and last codons could be matched. The gene and mRNA categories are the same in the MetaEuk GFF. The exon and CDS coordinates will be the same unless a small target overlap was allowed, due to which, the MetaEuk exon was shortened (see above). In this case, the CDS will report the shortening. In the sixth column you can find their individual bitsocres. The contig index starts at 1 and the start coordinate is always smaller than the end coordinate, as required by GFF. The last column contains the **TCS** identifier, followed by the low_coord of the prediction to support searching for sub-optimal exon sets (see section). Here is an example where a MetaEuk header of two exons is reported in GFF format:
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