/*
* Jalview - A Sequence Alignment Editor and Viewer ($$Version-Rel$$)
* Copyright (C) $$Year-Rel$$ The Jalview Authors
*
* This file is part of Jalview.
*
* Jalview is free software: you can redistribute it and/or
* modify it under the terms of the GNU General Public License
* as published by the Free Software Foundation, either version 3
* of the License, or (at your option) any later version.
*
* Jalview is distributed in the hope that it will be useful, but
* WITHOUT ANY WARRANTY; without even the implied warranty
* of MERCHANTABILITY or FITNESS FOR A PARTICULAR
* PURPOSE. See the GNU General Public License for more details.
*
* You should have received a copy of the GNU General Public License
* along with Jalview. If not, see .
* The Jalview Authors are detailed in the 'AUTHORS' file.
*/
package jalview.ext.ensembl;
import jalview.analysis.AlignmentUtils;
import jalview.analysis.Dna;
import jalview.bin.Cache;
import jalview.datamodel.Alignment;
import jalview.datamodel.AlignmentI;
import jalview.datamodel.DBRefEntry;
import jalview.datamodel.DBRefSource;
import jalview.datamodel.Mapping;
import jalview.datamodel.SequenceFeature;
import jalview.datamodel.SequenceI;
import jalview.exceptions.JalviewException;
import jalview.io.FastaFile;
import jalview.io.FileParse;
import jalview.io.gff.SequenceOntologyFactory;
import jalview.io.gff.SequenceOntologyI;
import jalview.util.Comparison;
import jalview.util.DBRefUtils;
import jalview.util.MapList;
import java.io.IOException;
import java.net.MalformedURLException;
import java.net.URL;
import java.util.ArrayList;
import java.util.Arrays;
import java.util.Collections;
import java.util.Comparator;
import java.util.List;
/**
* Base class for Ensembl sequence fetchers
*
* @see http://rest.ensembl.org/documentation/info/sequence_id
* @author gmcarstairs
*/
public abstract class EnsemblSeqProxy extends EnsemblRestClient
{
private static final String ALLELES = "alleles";
protected static final String PARENT = "Parent";
protected static final String ID = "ID";
protected static final String NAME = "Name";
protected static final String DESCRIPTION = "description";
/*
* enum for 'type' parameter to the /sequence REST service
*/
public enum EnsemblSeqType
{
/**
* type=genomic to fetch full dna including introns
*/
GENOMIC("genomic"),
/**
* type=cdna to fetch coding dna including UTRs
*/
CDNA("cdna"),
/**
* type=cds to fetch coding dna excluding UTRs
*/
CDS("cds"),
/**
* type=protein to fetch peptide product sequence
*/
PROTEIN("protein");
/*
* the value of the 'type' parameter to fetch this version of
* an Ensembl sequence
*/
private String type;
EnsemblSeqType(String t)
{
type = t;
}
public String getType()
{
return type;
}
}
/**
* A comparator to sort ranges into ascending start position order
*/
private class RangeSorter implements Comparator
{
boolean forwards;
RangeSorter(boolean forward)
{
forwards = forward;
}
@Override
public int compare(int[] o1, int[] o2)
{
return (forwards ? 1 : -1) * Integer.compare(o1[0], o2[0]);
}
}
/**
* Default constructor (to use rest.ensembl.org)
*/
public EnsemblSeqProxy()
{
super();
}
/**
* Constructor given the target domain to fetch data from
*/
public EnsemblSeqProxy(String d)
{
super(d);
}
/**
* Makes the sequence queries to Ensembl's REST service and returns an
* alignment consisting of the returned sequences.
*/
@Override
public AlignmentI getSequenceRecords(String query) throws Exception
{
// TODO use a String... query vararg instead?
// danger: accession separator used as a regex here, a string elsewhere
// in this case it is ok (it is just a space), but (e.g.) '\' would not be
List allIds = Arrays.asList(query
.split(getAccessionSeparator()));
AlignmentI alignment = null;
inProgress = true;
/*
* execute queries, if necessary in batches of the
* maximum allowed number of ids
*/
int maxQueryCount = getMaximumQueryCount();
for (int v = 0, vSize = allIds.size(); v < vSize; v += maxQueryCount)
{
int p = Math.min(vSize, v + maxQueryCount);
List ids = allIds.subList(v, p);
try
{
alignment = fetchSequences(ids, alignment);
} catch (Throwable r)
{
inProgress = false;
String msg = "Aborting ID retrieval after " + v
+ " chunks. Unexpected problem (" + r.getLocalizedMessage()
+ ")";
System.err.println(msg);
r.printStackTrace();
break;
}
}
if (alignment == null)
{
return null;
}
/*
* fetch and transfer genomic sequence features,
* fetch protein product and add as cross-reference
*/
for (String accId : allIds)
{
addFeaturesAndProduct(accId, alignment);
}
for (SequenceI seq : alignment.getSequences())
{
getCrossReferences(seq);
}
return alignment;
}
/**
* Fetches Ensembl features using the /overlap REST endpoint, and adds them to
* the sequence in the alignment. Also fetches the protein product, maps it
* from the CDS features of the sequence, and saves it as a cross-reference of
* the dna sequence.
*
* @param accId
* @param alignment
*/
protected void addFeaturesAndProduct(String accId, AlignmentI alignment)
{
if (alignment == null)
{
return;
}
try
{
/*
* get 'dummy' genomic sequence with exon, cds and variation features
*/
SequenceI genomicSequence = null;
EnsemblFeatures gffFetcher = new EnsemblFeatures(getDomain());
EnsemblFeatureType[] features = getFeaturesToFetch();
AlignmentI geneFeatures = gffFetcher.getSequenceRecords(accId,
features);
if (geneFeatures.getHeight() > 0)
{
genomicSequence = geneFeatures.getSequenceAt(0);
}
if (genomicSequence != null)
{
/*
* transfer features to the query sequence
*/
SequenceI querySeq = alignment.findName(accId);
if (transferFeatures(accId, genomicSequence, querySeq))
{
/*
* fetch and map protein product, and add it as a cross-reference
* of the retrieved sequence
*/
addProteinProduct(querySeq);
}
}
} catch (IOException e)
{
System.err.println("Error transferring Ensembl features: "
+ e.getMessage());
}
}
/**
* Returns those sequence feature types to fetch from Ensembl. We may want
* features either because they are of interest to the user, or as means to
* identify the locations of the sequence on the genomic sequence (CDS
* features identify CDS, exon features identify cDNA etc).
*
* @return
*/
protected abstract EnsemblFeatureType[] getFeaturesToFetch();
/**
* Fetches and maps the protein product, and adds it as a cross-reference of
* the retrieved sequence
*/
protected void addProteinProduct(SequenceI querySeq)
{
String accId = querySeq.getName();
try
{
AlignmentI protein = new EnsemblProtein(getDomain())
.getSequenceRecords(accId);
if (protein == null || protein.getHeight() == 0)
{
System.out.println("No protein product found for " + accId);
return;
}
SequenceI proteinSeq = protein.getSequenceAt(0);
/*
* need dataset sequences (to be the subject of mappings)
*/
proteinSeq.createDatasetSequence();
querySeq.createDatasetSequence();
MapList mapList = AlignmentUtils
.mapCdsToProtein(querySeq, proteinSeq);
if (mapList != null)
{
// clunky: ensure Uniprot xref if we have one is on mapped sequence
SequenceI ds = proteinSeq.getDatasetSequence();
// TODO: Verify ensp primary ref is on proteinSeq.getDatasetSequence()
Mapping map = new Mapping(ds, mapList);
DBRefEntry dbr = new DBRefEntry(getDbSource(),
getEnsemblDataVersion(), proteinSeq.getName(), map);
querySeq.getDatasetSequence().addDBRef(dbr);
DBRefEntry[] uprots = DBRefUtils.selectRefs(ds.getDBRefs(),
new String[] { DBRefSource.UNIPROT });
DBRefEntry[] upxrefs = DBRefUtils.selectRefs(querySeq.getDBRefs(),
new String[] { DBRefSource.UNIPROT });
if (uprots != null)
{
for (DBRefEntry up : uprots)
{
// locate local uniprot ref and map
List upx = DBRefUtils.searchRefs(upxrefs,
up.getAccessionId());
DBRefEntry upxref;
if (upx.size() != 0)
{
upxref = upx.get(0);
if (upx.size() > 1)
{
Cache.log
.warn("Implementation issue - multiple uniprot acc on product sequence.");
}
}
else
{
upxref = new DBRefEntry(DBRefSource.UNIPROT,
getEnsemblDataVersion(), up.getAccessionId());
}
Mapping newMap = new Mapping(ds, mapList);
upxref.setVersion(getEnsemblDataVersion());
upxref.setMap(newMap);
if (upx.size() == 0)
{
// add the new uniprot ref
querySeq.getDatasetSequence().addDBRef(upxref);
}
}
}
/*
* copy exon features to protein, compute peptide variants from dna
* variants and add as features on the protein sequence ta-da
*/
AlignmentUtils
.computeProteinFeatures(querySeq, proteinSeq, mapList);
}
} catch (Exception e)
{
System.err
.println(String.format("Error retrieving protein for %s: %s",
accId, e.getMessage()));
}
}
/**
* Get database xrefs from Ensembl, and attach them to the sequence
*
* @param seq
*/
protected void getCrossReferences(SequenceI seq)
{
while (seq.getDatasetSequence() != null)
{
seq = seq.getDatasetSequence();
}
EnsemblXref xrefFetcher = new EnsemblXref(getDomain(), getDbSource(),
getEnsemblDataVersion());
List xrefs = xrefFetcher.getCrossReferences(seq.getName());
for (DBRefEntry xref : xrefs)
{
seq.addDBRef(xref);
}
/*
* and add a reference to itself
*/
DBRefEntry self = new DBRefEntry(getDbSource(),
getEnsemblDataVersion(), seq.getName());
seq.addDBRef(self);
}
/**
* Fetches sequences for the list of accession ids and adds them to the
* alignment. Returns the extended (or created) alignment.
*
* @param ids
* @param alignment
* @return
* @throws JalviewException
* @throws IOException
*/
protected AlignmentI fetchSequences(List ids, AlignmentI alignment)
throws JalviewException, IOException
{
if (!isEnsemblAvailable())
{
inProgress = false;
throw new JalviewException("ENSEMBL Rest API not available.");
}
FileParse fp = getSequenceReader(ids);
if (fp == null)
{
return alignment;
}
FastaFile fr = new FastaFile(fp);
if (fr.hasWarningMessage())
{
System.out.println(String.format(
"Warning when retrieving %d ids %s\n%s", ids.size(),
ids.toString(), fr.getWarningMessage()));
}
else if (fr.getSeqs().size() != ids.size())
{
System.out.println(String.format(
"Only retrieved %d sequences for %d query strings", fr
.getSeqs().size(), ids.size()));
}
if (fr.getSeqs().size() == 1 && fr.getSeqs().get(0).getLength() == 0)
{
/*
* POST request has returned an empty FASTA file e.g. for invalid id
*/
throw new IOException("No data returned for " + ids);
}
if (fr.getSeqs().size() > 0)
{
AlignmentI seqal = new Alignment(fr.getSeqsAsArray());
for (SequenceI sq : seqal.getSequences())
{
if (sq.getDescription() == null)
{
sq.setDescription(getDbName());
}
String name = sq.getName();
if (ids.contains(name)
|| ids.contains(name.replace("ENSP", "ENST")))
{
DBRefEntry dbref = DBRefUtils.parseToDbRef(sq, getDbSource(),
getEnsemblDataVersion(), name);
sq.addDBRef(dbref);
}
}
if (alignment == null)
{
alignment = seqal;
}
else
{
alignment.append(seqal);
}
}
return alignment;
}
/**
* Returns the URL for the REST call
*
* @return
* @throws MalformedURLException
*/
@Override
protected URL getUrl(List ids) throws MalformedURLException
{
/*
* a single id is included in the URL path
* multiple ids go in the POST body instead
*/
StringBuffer urlstring = new StringBuffer(128);
urlstring.append(getDomain() + "/sequence/id");
if (ids.size() == 1)
{
urlstring.append("/").append(ids.get(0));
}
// @see https://github.com/Ensembl/ensembl-rest/wiki/Output-formats
urlstring.append("?type=").append(getSourceEnsemblType().getType());
urlstring.append(("&Accept=text/x-fasta"));
URL url = new URL(urlstring.toString());
return url;
}
/**
* A sequence/id POST request currently allows up to 50 queries
*
* @see http://rest.ensembl.org/documentation/info/sequence_id_post
*/
@Override
public int getMaximumQueryCount()
{
return 50;
}
@Override
protected boolean useGetRequest()
{
return false;
}
@Override
protected String getRequestMimeType(boolean multipleIds)
{
return multipleIds ? "application/json" : "text/x-fasta";
}
@Override
protected String getResponseMimeType()
{
return "text/x-fasta";
}
/**
*
* @return the configured sequence return type for this source
*/
protected abstract EnsemblSeqType getSourceEnsemblType();
/**
* Returns a list of [start, end] genomic ranges corresponding to the sequence
* being retrieved.
*
* The correspondence between the frames of reference is made by locating
* those features on the genomic sequence which identify the retrieved
* sequence. Specifically
*
* - genomic sequence is identified by "transcript" features with
* ID=transcript:transcriptId
* - cdna sequence is identified by "exon" features with
* Parent=transcript:transcriptId
* - cds sequence is identified by "CDS" features with
* Parent=transcript:transcriptId
*
*
* The returned ranges are sorted to run forwards (for positive strand) or
* backwards (for negative strand). Aborts and returns null if both positive
* and negative strand are found (this should not normally happen).
*
* @param sourceSequence
* @param accId
* @param start
* the start position of the sequence we are mapping to
* @return
*/
protected MapList getGenomicRangesFromFeatures(SequenceI sourceSequence,
String accId, int start)
{
SequenceFeature[] sfs = sourceSequence.getSequenceFeatures();
if (sfs == null)
{
return null;
}
/*
* generously initial size for number of cds regions
* (worst case titin Q8WZ42 has c. 313 exons)
*/
List regions = new ArrayList(100);
int mappedLength = 0;
int direction = 1; // forward
boolean directionSet = false;
for (SequenceFeature sf : sfs)
{
/*
* accept the target feature type or a specialisation of it
* (e.g. coding_exon for exon)
*/
if (identifiesSequence(sf, accId))
{
int strand = sf.getStrand();
strand = strand == 0 ? 1 : strand; // treat unknown as forward
if (directionSet && strand != direction)
{
// abort - mix of forward and backward
System.err.println("Error: forward and backward strand for "
+ accId);
return null;
}
direction = strand;
directionSet = true;
/*
* add to CDS ranges, semi-sorted forwards/backwards
*/
if (strand < 0)
{
regions.add(0, new int[] { sf.getEnd(), sf.getBegin() });
}
else
{
regions.add(new int[] { sf.getBegin(), sf.getEnd() });
}
mappedLength += Math.abs(sf.getEnd() - sf.getBegin() + 1);
if (!isSpliceable())
{
/*
* 'gene' sequence is contiguous so we can stop as soon as its
* identifying feature has been found
*/
break;
}
}
}
if (regions.isEmpty())
{
System.out.println("Failed to identify target sequence for " + accId
+ " from genomic features");
return null;
}
/*
* a final sort is needed since Ensembl returns CDS sorted within source
* (havana / ensembl_havana)
*/
Collections.sort(regions, new RangeSorter(direction == 1));
List to = Arrays.asList(new int[] { start,
start + mappedLength - 1 });
return new MapList(regions, to, 1, 1);
}
/**
* Answers true if the sequence being retrieved may occupy discontiguous
* regions on the genomic sequence.
*/
protected boolean isSpliceable()
{
return true;
}
/**
* Returns true if the sequence feature marks positions of the genomic
* sequence feature which are within the sequence being retrieved. For
* example, an 'exon' feature whose parent is the target transcript marks the
* cdna positions of the transcript.
*
* @param sf
* @param accId
* @return
*/
protected abstract boolean identifiesSequence(SequenceFeature sf,
String accId);
/**
* Transfers the sequence feature to the target sequence, locating its start
* and end range based on the mapping. Features which do not overlap the
* target sequence are ignored.
*
* @param sf
* @param targetSequence
* @param mapping
* mapping from the sequence feature's coordinates to the target
* sequence
* @param forwardStrand
*/
protected void transferFeature(SequenceFeature sf,
SequenceI targetSequence, MapList mapping, boolean forwardStrand)
{
int start = sf.getBegin();
int end = sf.getEnd();
int[] mappedRange = mapping.locateInTo(start, end);
if (mappedRange != null)
{
SequenceFeature copy = new SequenceFeature(sf);
copy.setBegin(Math.min(mappedRange[0], mappedRange[1]));
copy.setEnd(Math.max(mappedRange[0], mappedRange[1]));
if (".".equals(copy.getFeatureGroup()))
{
copy.setFeatureGroup(getDbSource());
}
targetSequence.addSequenceFeature(copy);
/*
* for sequence_variant on reverse strand, have to convert the allele
* values to their complements
*/
if (!forwardStrand
&& SequenceOntologyFactory.getInstance().isA(sf.getType(),
SequenceOntologyI.SEQUENCE_VARIANT))
{
reverseComplementAlleles(copy);
}
}
}
/**
* Change the 'alleles' value of a feature by converting to complementary
* bases, and also update the feature description to match
*
* @param sf
*/
static void reverseComplementAlleles(SequenceFeature sf)
{
final String alleles = (String) sf.getValue(ALLELES);
if (alleles == null)
{
return;
}
StringBuilder complement = new StringBuilder(alleles.length());
for (String allele : alleles.split(","))
{
reverseComplementAllele(complement, allele);
}
String comp = complement.toString();
sf.setValue(ALLELES, comp);
sf.setDescription(comp);
/*
* replace value of "alleles=" in sf.ATTRIBUTES as well
* so 'output as GFF' shows reverse complement alleles
*/
String atts = sf.getAttributes();
if (atts != null)
{
atts = atts.replace(ALLELES + "=" + alleles, ALLELES + "=" + comp);
sf.setAttributes(atts);
}
}
/**
* Makes the 'reverse complement' of the given allele and appends it to the
* buffer, after a comma separator if not the first
*
* @param complement
* @param allele
*/
static void reverseComplementAllele(StringBuilder complement,
String allele)
{
if (complement.length() > 0)
{
complement.append(",");
}
/*
* some 'alleles' are actually descriptive terms
* e.g. HGMD_MUTATION, PhenCode_variation
* - we don't want to 'reverse complement' these
*/
if (!Comparison.isNucleotideSequence(allele, true))
{
complement.append(allele);
}
else
{
for (int i = allele.length() - 1; i >= 0; i--)
{
complement.append(Dna.getComplement(allele.charAt(i)));
}
}
}
/**
* Transfers features from sourceSequence to targetSequence
*
* @param accessionId
* @param sourceSequence
* @param targetSequence
* @return true if any features were transferred, else false
*/
protected boolean transferFeatures(String accessionId,
SequenceI sourceSequence, SequenceI targetSequence)
{
if (sourceSequence == null || targetSequence == null)
{
return false;
}
// long start = System.currentTimeMillis();
SequenceFeature[] sfs = sourceSequence.getSequenceFeatures();
MapList mapping = getGenomicRangesFromFeatures(sourceSequence,
accessionId, targetSequence.getStart());
if (mapping == null)
{
return false;
}
boolean result = transferFeatures(sfs, targetSequence, mapping,
accessionId);
// System.out.println("transferFeatures (" + (sfs.length) + " --> "
// + targetSequence.getSequenceFeatures().length + ") to "
// + targetSequence.getName()
// + " took " + (System.currentTimeMillis() - start) + "ms");
return result;
}
/**
* Transfer features to the target sequence. The start/end positions are
* converted using the mapping. Features which do not overlap are ignored.
* Features whose parent is not the specified identifier are also ignored.
*
* @param features
* @param targetSequence
* @param mapping
* @param parentId
* @return
*/
protected boolean transferFeatures(SequenceFeature[] features,
SequenceI targetSequence, MapList mapping, String parentId)
{
final boolean forwardStrand = mapping.isFromForwardStrand();
/*
* sort features by start position (which corresponds to end
* position descending if reverse strand) so as to add them in
* 'forwards' order to the target sequence
*/
sortFeatures(features, forwardStrand);
boolean transferred = false;
for (SequenceFeature sf : features)
{
if (retainFeature(sf, parentId))
{
transferFeature(sf, targetSequence, mapping, forwardStrand);
transferred = true;
}
}
return transferred;
}
/**
* Sort features by start position ascending (if on forward strand), or end
* position descending (if on reverse strand)
*
* @param features
* @param forwardStrand
*/
protected static void sortFeatures(SequenceFeature[] features,
final boolean forwardStrand)
{
Arrays.sort(features, new Comparator()
{
@Override
public int compare(SequenceFeature o1, SequenceFeature o2)
{
if (forwardStrand)
{
return Integer.compare(o1.getBegin(), o2.getBegin());
}
else
{
return Integer.compare(o2.getEnd(), o1.getEnd());
}
}
});
}
/**
* Answers true if the feature type is one we want to keep for the sequence.
* Some features are only retrieved in order to identify the sequence range,
* and may then be discarded as redundant information (e.g. "CDS" feature for
* a CDS sequence).
*/
@SuppressWarnings("unused")
protected boolean retainFeature(SequenceFeature sf, String accessionId)
{
return true; // override as required
}
/**
* Answers true if the feature has a Parent which refers to the given
* accession id, or if the feature has no parent. Answers false if the
* feature's Parent is for a different accession id.
*
* @param sf
* @param identifier
* @return
*/
protected boolean featureMayBelong(SequenceFeature sf, String identifier)
{
String parent = (String) sf.getValue(PARENT);
// using contains to allow for prefix "gene:", "transcript:" etc
if (parent != null && !parent.contains(identifier))
{
// this genomic feature belongs to a different transcript
return false;
}
return true;
}
@Override
public String getDescription()
{
return "Ensembl " + getSourceEnsemblType().getType()
+ " sequence with variant features";
}
/**
* Returns a (possibly empty) list of features on the sequence which have the
* specified sequence ontology type (or a sub-type of it), and the given
* identifier as parent
*
* @param sequence
* @param type
* @param parentId
* @return
*/
protected List findFeatures(SequenceI sequence,
String type, String parentId)
{
List result = new ArrayList();
SequenceFeature[] sfs = sequence.getSequenceFeatures();
if (sfs != null)
{
SequenceOntologyI so = SequenceOntologyFactory.getInstance();
for (SequenceFeature sf : sfs)
{
if (so.isA(sf.getType(), type))
{
String parent = (String) sf.getValue(PARENT);
if (parent.equals(parentId))
{
result.add(sf);
}
}
}
}
return result;
}
/**
* Answers true if the feature type is either 'NMD_transcript_variant' or
* 'transcript' or one of its sub-types in the Sequence Ontology. This is
* needed because NMD_transcript_variant behaves like 'transcript' in Ensembl
* although strictly speaking it is not (it is a sub-type of
* sequence_variant).
*
* @param featureType
* @return
*/
public static boolean isTranscript(String featureType)
{
return SequenceOntologyI.NMD_TRANSCRIPT_VARIANT.equals(featureType)
|| SequenceOntologyFactory.getInstance().isA(featureType,
SequenceOntologyI.TRANSCRIPT);
}
}