void driver(const vector<FILE*>& splice_coords_files,
const vector<FILE*>& insertion_coords_files,
const vector<FILE*>& deletion_coords_files,
ifstream& ref_stream)
{
char splice_buf[2048];
RefSequenceTable rt(true);
JunctionSet junctions;
for (size_t i = 0; i < splice_coords_files.size(); ++i)
{
FILE* splice_coords = splice_coords_files[i];
if (!splice_coords)
continue;
while (fgets(splice_buf, 2048, splice_coords))
{
char* nl = strrchr(splice_buf, '\n');
char* buf = splice_buf;
if (nl) *nl = 0;
/**
Fields are:
1) reference name
2) left coord of splice (last char of the left exon)
3) right coord of splice (first char of the right exon)
*/
char* ref_name = get_token((char**)&buf, "\t");
char* scan_left_coord = get_token((char**)&buf, "\t");
char* scan_right_coord = get_token((char**)&buf, "\t");
char* orientation = get_token((char**)&buf, "\t");
if (!scan_left_coord || !scan_right_coord || !orientation)
{
fprintf(stderr,"Error: malformed splice coordinate record\n");
exit(1);
}
uint32_t ref_id = rt.get_id(ref_name, NULL, 0);
uint32_t left_coord = atoi(scan_left_coord);
uint32_t right_coord = atoi(scan_right_coord);
bool antisense = *orientation == '-';
junctions.insert(make_pair<Junction, JunctionStats>(Junction(ref_id, left_coord, right_coord, antisense), JunctionStats()));
}
}
/*
* Read in the deletion coordinates
* and store in a set
*/
std::set<Deletion> deletions;
for(size_t i=0; i < deletion_coords_files.size(); ++i){
FILE* deletion_coords = deletion_coords_files[i];
if(!deletion_coords){
continue;
}
while (fgets(splice_buf, 2048, deletion_coords))
{
char* nl = strrchr(splice_buf, '\n');
char* buf = splice_buf;
if (nl) *nl = 0;
/**
Fields are:
1) reference name
2) left coord of splice (last char of the left exon)
3) right coord of splice (first char of the right exon)
*/
char* ref_name = get_token((char**)&buf, "\t");
char* scan_left_coord = get_token((char**)&buf, "\t");
char* scan_right_coord = get_token((char**)&buf, "\t");
if (!scan_left_coord || !scan_right_coord)
{
fprintf(stderr,"Error: malformed deletion coordinate record\n");
exit(1);
}
/*
* Note that when reading in a deletion, the left co-ord is the position of the
* first deleted based. Since we are co-opting the junction data structure, need
* to fix up this location
*/
uint32_t ref_id = rt.get_id(ref_name, NULL, 0);
uint32_t left_coord = atoi(scan_left_coord);
uint32_t right_coord = atoi(scan_right_coord);
deletions.insert(Deletion(ref_id, left_coord - 1, right_coord, false));
}
}
/*
* Read in the insertion coordinates
* and store in a set
*/
std::set<Insertion> insertions;
for(size_t i=0; i < insertion_coords_files.size(); ++i){
FILE* insertion_coords = insertion_coords_files[i];
if(!insertion_coords){
continue;
}
while(fgets(splice_buf, 2048, insertion_coords)){
char* nl = strrchr(splice_buf, '\n');
char* buf = splice_buf;
if (nl) *nl = 0;
char* ref_name = get_token((char**)&buf, "\t");
char* scan_left_coord = get_token((char**)&buf, "\t");
char* scan_right_coord = get_token((char**)&buf, "\t");
char* scan_sequence = get_token((char**)&buf, "\t");
if (!scan_left_coord || !scan_sequence || !scan_right_coord)
{
fprintf(stderr,"Error: malformed insertion coordinate record\n");
exit(1);
}
seqan::Dna5String sequence = seqan::Dna5String(scan_sequence);
bool containsN = false;
for(size_t index = 0; index < seqan::length(sequence); index += 1){
/*
* Don't allow any ambiguities in the insertion
*/
if(sequence[index] == 'N'){
containsN = true;
break;
}
}
if(containsN){
continue;
}
seqan::CharString charSequence = sequence;
uint32_t ref_id = rt.get_id(ref_name,NULL,0);
uint32_t left_coord = atoi(scan_left_coord);
insertions.insert(Insertion(ref_id, left_coord, seqan::toCString(charSequence)));
}
}
typedef RefSequenceTable::Sequence Reference;
while(ref_stream.good() &&
!ref_stream.eof())
{
Reference ref_str;
string name;
readMeta(ref_stream, name, Fasta());
string::size_type space_pos = name.find_first_of(" \t\r");
if (space_pos != string::npos)
{
name.resize(space_pos);
}
read(ref_stream, ref_str, Fasta());
uint32_t refid = rt.get_id(name, NULL, 0);
Junction dummy_left(refid, 0, 0, true);
Junction dummy_right(refid, VMAXINT32, VMAXINT32, true);
pair<JunctionSet::iterator, JunctionSet::iterator> r;
r.first = junctions.lower_bound(dummy_left);
r.second = junctions.upper_bound(dummy_right);
JunctionSet::iterator itr = r.first;
while(itr != r.second && itr != junctions.end())
{
print_splice(itr->first, read_length, itr->first.antisense ? "GTAG|rev" : "GTAG|fwd", ref_str, name, cout);
++itr;
}
}
ref_stream.clear();
ref_stream.seekg(0, ios::beg);
while(ref_stream.good() &&
!ref_stream.eof())
{
Reference ref_str;
string name;
readMeta(ref_stream, name, Fasta());
string::size_type space_pos = name.find_first_of(" \t\r");
if (space_pos != string::npos)
{
name.resize(space_pos);
}
read(ref_stream, ref_str, Fasta());
uint32_t refid = rt.get_id(name, NULL,0);
Deletion dummy_left(refid, 0, 0, true);
Deletion dummy_right(refid, VMAXINT32, VMAXINT32, true);
pair<std::set<Deletion>::iterator, std::set<Deletion>::iterator> r;
r.first = deletions.lower_bound(dummy_left);
r.second = deletions.upper_bound(dummy_right);
std::set<Deletion>::iterator itr = r.first;
while(itr != r.second && itr != deletions.end())
{
print_splice((Junction)*itr, read_length, itr->antisense ? "del|rev" : "del|fwd", ref_str, name, cout);
++itr;
}
}
ref_stream.clear();
ref_stream.seekg(0, ios::beg);
while(ref_stream.good() &&
!ref_stream.eof())
{
Reference ref_str;
string name;
readMeta(ref_stream, name, Fasta());
string::size_type space_pos = name.find_first_of(" \t\r");
if (space_pos != string::npos)
{
name.resize(space_pos);
}
read(ref_stream, ref_str, Fasta());
uint32_t refid = rt.get_id(name, NULL,0);
Insertion dummy_left(refid, 0, "");
Insertion dummy_right(refid, VMAXINT32, "");
std::set<Insertion>::iterator itr = insertions.lower_bound(dummy_left);
std::set<Insertion>::iterator upper = insertions.upper_bound(dummy_right);
while(itr != upper && itr != insertions.end()){
print_insertion(*itr, read_length, ref_str, name, cout);
++itr;
}
}
}
void knockout_shadow_junctions(JunctionSet& junctions)
{
vector<uint32_t> ref_ids;
for (JunctionSet::iterator i = junctions.begin(); i != junctions.end(); ++i)
{
ref_ids.push_back(i->first.refid);
}
sort(ref_ids.begin(), ref_ids.end());
vector<uint32_t>::iterator new_end = unique(ref_ids.begin(), ref_ids.end());
ref_ids.erase(new_end, ref_ids.end());
for(size_t i = 0; i < ref_ids.size(); ++i)
{
uint32_t refid = ref_ids[i];
Junction dummy_left(refid, 0, 0, true);
Junction dummy_right(refid, 0xFFFFFFFF, 0xFFFFFFFF, true);
pair<JunctionSet::iterator, JunctionSet::iterator> r;
r.first = junctions.lower_bound(dummy_left);
r.second = junctions.upper_bound(dummy_right);
JunctionSet::iterator itr = r.first;
while(itr != r.second && itr != junctions.end())
{
if (itr->second.accepted)
{
Junction fuzzy_left = itr->first;
Junction fuzzy_right = itr->first;
fuzzy_left.left -= min_anchor_len;
fuzzy_right.right += min_anchor_len;
fuzzy_left.antisense = !itr->first.antisense;
fuzzy_right.antisense = !itr->first.antisense;
pair<JunctionSet::iterator, JunctionSet::iterator> s;
s.first = junctions.lower_bound(fuzzy_left);
s.second = junctions.upper_bound(fuzzy_right);
JunctionSet::iterator itr2 = s.first;
int junc_support = itr->second.supporting_hits;
while(itr2 != s.second && itr2 != junctions.end())
{
int left_diff = itr->first.left - itr2->first.left;
int right_diff = itr->first.right - itr2->first.right;
if (itr != itr2 &&
itr->first.antisense != itr2->first.antisense &&
(left_diff < min_anchor_len || right_diff < min_anchor_len))
{
if (junc_support < itr2->second.supporting_hits)
itr->second.accepted = false;
}
++itr2;
}
}
++itr;
}
}
}