annotate tools/protein_analysis/rxlr_motifs.py @ 18:eb6ac44d4b8e draft

Suite v0.2.8, record Promoter 2 verion + misc internal updates
author peterjc
date Tue, 01 Sep 2015 09:56:36 -0400
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1 #!/usr/bin/env python
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2 """Implements assorted RXLR motif methods from the literature
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3
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4 This script takes exactly four command line arguments:
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5 * Protein FASTA filename
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6 * Number of threads
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7 * Model name (Bhattacharjee2006, Win2007, Whisson2007)
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8 * Output tabular filename
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9
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10 The model names are:
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11
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12 Bhattacharjee2006: Simple regular expression search for RXLR
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13 with additional requirements for positioning and signal peptide.
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14
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15 Win2007: Simple regular expression search for RXLR, but with
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16 different positional requirements.
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17
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18 Whisson2007: As Bhattacharjee2006 but with a more complex regular
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19 expression to look for RXLR-EER domain, and additionally calls HMMER.
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20
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21 See the help text in the accompanying Galaxy tool XML file for more
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22 details including the full references.
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23
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24 Note:
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25
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26 Bhattacharjee et al. (2006) and Win et al. (2007) used SignalP v2.0,
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27 which is no longer available. The current release is SignalP v3.0
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28 (Mar 5, 2007). We have therefore opted to use the NN Ymax position for
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29 the predicted cleavage site, as this is expected to be more accurate.
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30 Also note that the HMM score values have changed from v2.0 to v3.0.
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31 Whisson et al. (2007) used SignalP v3.0 anyway.
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32
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33 Whisson et al. (2007) used HMMER 2.3.2, and althought their HMM model
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34 can still be used with hmmsearch from HMMER 3 this this does give
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35 slightly different results. We expect the hmmsearch from HMMER 2.3.2
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36 (the last stable release of HMMER 2) to be present on the path under
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37 the name hmmsearch2 (allowing it to co-exist with HMMER 3).
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38 """
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39 import os
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40 import sys
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41 import re
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42 import subprocess
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43 from seq_analysis_utils import sys_exit, fasta_iterator
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44
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45 if "-v" in sys.argv:
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46 print("RXLR Motifs v0.0.10")
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47 sys.exit(0)
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48
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49 if len(sys.argv) != 5:
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50 sys_exit("Requires four arguments: protein FASTA filename, threads, model, and output filename")
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52 fasta_file, threads, model, tabular_file = sys.argv[1:]
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53 hmm_output_file = tabular_file + ".hmm.tmp"
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54 signalp_input_file = tabular_file + ".fasta.tmp"
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55 signalp_output_file = tabular_file + ".tabular.tmp"
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56 min_signalp_hmm = 0.9
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57 hmmer_search = "hmmsearch2"
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58
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59 if model == "Bhattacharjee2006":
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60 signalp_trunc = 70
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61 re_rxlr = re.compile("R.LR")
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62 min_sp = 10
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63 max_sp = 40
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64 max_sp_rxlr = 100
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65 min_rxlr_start = 1
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66 # Allow signal peptide to be at most 40aa, and want RXLR to be
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67 # within 100aa, therefore for the prescreen the max start is 140:
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68 max_rxlr_start = max_sp + max_sp_rxlr
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69 elif model == "Win2007":
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70 signalp_trunc = 70
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71 re_rxlr = re.compile("R.LR")
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72 min_sp = 10
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73 max_sp = 40
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74 min_rxlr_start = 30
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75 max_rxlr_start = 60
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76 # No explicit limit on separation of signal peptide clevage
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77 # and RXLR, but shortest signal peptide is 10, and furthest
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78 # away RXLR is 60, so effectively limit is 50.
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79 max_sp_rxlr = max_rxlr_start - min_sp + 1
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80 elif model == "Whisson2007":
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81 signalp_trunc = 0 # zero for no truncation
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82 re_rxlr = re.compile("R.LR.{,40}[ED][ED][KR]")
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83 min_sp = 10
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84 max_sp = 40
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85 max_sp_rxlr = 100
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86 min_rxlr_start = 1
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87 max_rxlr_start = max_sp + max_sp_rxlr
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88 else:
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89 sys_exit("Did not recognise the model name %r\n"
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90 "Use Bhattacharjee2006, Win2007, or Whisson2007" % model)
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91
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92
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93 def get_hmmer_version(exe, required=None):
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94 cmd = "%s -h" % exe
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95 try:
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96 child = subprocess.Popen([exe, "-h"], stdout=subprocess.PIPE, stderr=subprocess.PIPE)
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97 except OSError:
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98 raise ValueError("Could not run %s" % exe)
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99 stdout, stderr = child.communicate()
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100 if required:
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101 return required in stdout
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102 elif "HMMER 2" in stdout:
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103 return 2
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104 elif "HMMER 3" in stdout:
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105 return 3
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106 else:
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107 raise ValueError("Could not determine version of %s" % exe)
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108
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109
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110 #Run hmmsearch for Whisson et al. (2007)
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111 if model == "Whisson2007":
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112 hmm_file = os.path.join(os.path.split(sys.argv[0])[0],
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113 "whisson_et_al_rxlr_eer_cropped.hmm")
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114 if not os.path.isfile(hmm_file):
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115 sys_exit("Missing HMM file for Whisson et al. (2007)")
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116 if not get_hmmer_version(hmmer_search, "HMMER 2.3.2 (Oct 2003)"):
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117 sys_exit("Missing HMMER 2.3.2 (Oct 2003) binary, %s" % hmmer_search)
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118
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119 hmm_hits = set()
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120 valid_ids = set()
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121 for title, seq in fasta_iterator(fasta_file):
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122 name = title.split(None,1)[0]
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123 if name in valid_ids:
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124 sys_exit("Duplicated identifier %r" % name)
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125 else:
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126 valid_ids.add(name)
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127 if not valid_ids:
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128 # Special case, don't need to run HMMER if there are no sequences
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129 pass
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130 else:
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131 # I've left the code to handle HMMER 3 in situ, in case
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132 # we revisit the choice to insist on HMMER 2.
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133 hmmer3 = (3 == get_hmmer_version(hmmer_search))
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134 # Using zero (or 5.6?) for bitscore threshold
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135 if hmmer3:
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136 # The HMMER3 table output is easy to parse
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137 # In HMMER3 can't use both -T and -E
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138 cmd = "%s -T 0 --tblout %s --noali %s %s > /dev/null" \
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139 % (hmmer_search, hmm_output_file, hmm_file, fasta_file)
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140 else:
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141 # For HMMER2 we are stuck with parsing stdout
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142 # Put 1e6 to effectively have no expectation threshold (otherwise
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143 # HMMER defaults to 10 and the calculated e-value depends on the
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144 # input FASTA file, and we can loose hits of interest).
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145 cmd = "%s -T 0 -E 1e6 %s %s > %s" \
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146 % (hmmer_search, hmm_file, fasta_file, hmm_output_file)
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147 return_code = os.system(cmd)
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148 if return_code:
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149 sys_exit("Error %i from hmmsearch:\n%s" % (return_code, cmd), return_code)
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150
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151 handle = open(hmm_output_file)
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152 for line in handle:
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153 if not line.strip():
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154 # We expect blank lines in the HMMER2 stdout
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155 continue
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156 elif line.startswith("#"):
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157 # Header
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158 continue
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159 else:
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160 name = line.split(None,1)[0]
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161 #Should be a sequence name in the HMMER3 table output.
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162 #Could be anything in the HMMER2 stdout.
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163 if name in valid_ids:
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164 hmm_hits.add(name)
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165 elif hmmer3:
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166 sys_exit("Unexpected identifer %r in hmmsearch output" % name)
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167 handle.close()
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168 # if hmmer3:
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169 # print "HMMER3 hits for %i/%i" % (len(hmm_hits), len(valid_ids))
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170 # else:
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171 # print "HMMER2 hits for %i/%i" % (len(hmm_hits), len(valid_ids))
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172 # print "%i/%i matched HMM" % (len(hmm_hits), len(valid_ids))
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173 os.remove(hmm_output_file)
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174 del valid_ids
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175
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176
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177 # Prepare short list of candidates containing RXLR to pass to SignalP
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178 assert min_rxlr_start > 0, "Min value one, since zero based counting"
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179 count = 0
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180 total = 0
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181 handle = open(signalp_input_file, "w")
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182 for title, seq in fasta_iterator(fasta_file):
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183 total += 1
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184 name = title.split(None,1)[0]
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185 match = re_rxlr.search(seq[min_rxlr_start-1:].upper())
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186 if match and min_rxlr_start - 1 + match.start() + 1 <= max_rxlr_start:
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187 # This is a potential RXLR, depending on the SignalP results.
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188 # Might as well truncate the sequence now, makes the temp file smaller
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189 if signalp_trunc:
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190 handle.write(">%s (truncated)\n%s\n" % (name, seq[:signalp_trunc]))
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191 else:
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192 # Does it matter we don't line wrap?
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193 handle.write(">%s\n%s\n" % (name, seq))
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194 count += 1
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195 handle.close()
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196 # print "Running SignalP on %i/%i potentials." % (count, total)
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197
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198
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199 # Run SignalP (using our wrapper script to get multi-core support etc)
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200 signalp_script = os.path.join(os.path.split(sys.argv[0])[0], "signalp3.py")
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201 if not os.path.isfile(signalp_script):
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202 sys_exit("Error - missing signalp3.py script")
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203 cmd = "python %s euk %i %s %s %s" % (signalp_script, signalp_trunc, threads, signalp_input_file, signalp_output_file)
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204 return_code = os.system(cmd)
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205 if return_code:
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206 sys_exit("Error %i from SignalP:\n%s" % (return_code, cmd))
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207 # print "SignalP done"
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208
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209
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210 def parse_signalp(filename):
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211 """Parse SignalP output, yield tuples of ID, HMM_Sprob_score and NN predicted signal peptide length.
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212
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213 For signal peptide length we use NN_Ymax_pos (minus one).
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214 """
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215 handle = open(filename)
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216 line = handle.readline()
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217 assert line.startswith("#ID\t"), line
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218 for line in handle:
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219 parts = line.rstrip("\t").split("\t")
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220 assert len(parts)==20, repr(line)
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221 yield parts[0], float(parts[18]), int(parts[5])-1
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222 handle.close()
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223
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224
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225 # Parse SignalP results and apply the strict RXLR criteria
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226 total = 0
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227 tally = dict()
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228 handle = open(tabular_file, "w")
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229 handle.write("#ID\t%s\n" % model)
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230 signalp_results = parse_signalp(signalp_output_file)
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231 for title, seq in fasta_iterator(fasta_file):
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232 total += 1
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233 rxlr = "N"
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234 name = title.split(None,1)[0]
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235 match = re_rxlr.search(seq[min_rxlr_start-1:].upper())
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236 if match and min_rxlr_start - 1 + match.start() + 1 <= max_rxlr_start:
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237 del match
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238 # This was the criteria for calling SignalP,
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239 #so it will be in the SignalP results.
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240 sp_id, sp_hmm_score, sp_nn_len = signalp_results.next()
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241 assert name == sp_id, "%s vs %s" % (name, sp_id)
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242 if sp_hmm_score >= min_signalp_hmm and min_sp <= sp_nn_len <= max_sp:
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243 match = re_rxlr.search(seq[sp_nn_len:].upper())
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244 if match and match.start() + 1 <= max_sp_rxlr: # 1-based counting
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245 rxlr_start = sp_nn_len + match.start() + 1
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246 if min_rxlr_start <= rxlr_start <= max_rxlr_start:
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247 rxlr = "Y"
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248 if model == "Whisson2007":
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249 # Combine the signalp with regular expression heuristic and the HMM
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250 if name in hmm_hits and rxlr == "N":
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251 rxlr = "hmm" # HMM only
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252 elif rxlr == "N":
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253 rxlr = "neither" # Don't use N (no)
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254 elif name not in hmm_hits and rxlr == "Y":
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255 rxlr = "re" # Heuristic only
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256 # Now have a four way classifier: Y, hmm, re, neither
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257 # and count is the number of Y results (both HMM and heuristic)
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258 handle.write("%s\t%s\n" % (name, rxlr))
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259 try:
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260 tally[rxlr] += 1
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261 except KeyError:
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262 tally[rxlr] = 1
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263 handle.close()
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264 assert sum(tally.values()) == total
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265
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266 # Check the iterator is finished
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267 try:
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268 signalp_results.next()
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269 assert False, "Unexpected data in SignalP output"
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270 except StopIteration:
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271 pass
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272
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273 # Cleanup
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274 os.remove(signalp_input_file)
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275 os.remove(signalp_output_file)
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276
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277 # Short summary to stdout for Galaxy's info display
6
a290c6d4e658 Migrated tool version 0.0.9 from old tool shed archive to new tool shed repository
peterjc
parents:
diff changeset
278 print "%s for %i sequences:" % (model, total)
a290c6d4e658 Migrated tool version 0.0.9 from old tool shed archive to new tool shed repository
peterjc
parents:
diff changeset
279 print ", ".join("%s = %i" % kv for kv in sorted(tally.iteritems()))