Whole-genome motif discovery

1. Whole-genome motif discovery

2. 7 9 Challenges in Computational Biology 4 Genome Assembly Gene Finding Regulatory motif discovery DNA Sequence alignment Comparative Genomics TCATGCTAT TCGTGATAA TGAGGATAT TTATCATAT TTATGATTT Database lookup Evolutionary Theory RNA folding Gene expression analysis RNA transcript 10 Cluster discovery Gibbs sampling 12 Protein network analysis 13 Regulatory network inference 14 Emerging network properties

3. TTATATTGAATTTTCAAAAATTCTTACTTTTTTTTTGGATGGACGCAAAGAAGTTTAATAATCATATTACATGGCATTACCACCATATACATATCCATATCTAATCTTACTTATATGTTGTGGAAATGTAAAGAGCCCCATTATCTTAGCCTAAAAAAACCTTCTCTTTGGAACTTTCAGTAATACGCTTAACTGCTCATTGCTATATTGAAGTACGGATTAGAAGCCGCCGAGCGGGCGACAGCCCTCCGACGGAAGACTCTCCTCCGTGCGTCCTCGTCTTCACCGGTCGCGTTCCTGAAACGCAGATGTGCCTCGCGCCGCACTGCTCCGAACAATAAAGATTCTACAATACTAGCTTTTATGGTTATGAAGAGGAAAAATTGGCAGTAACCTGGCCCCACAAACCTTCAAATTAACGAATCAAATTAACAACCATAGGATGATAATGCGATTAGTTTTTTAGCCTTATTTCTGGGGTAATTAATCAGCGAAGCGATGATTTTTGATCTATTAACAGATATATAAATGGAAAAGCTGCATAACCACTTTAACTAATACTTTCAACATTTTCAGTTTGTATTACTTCTTATTCAAATGTCATAAAAGTATCAACAAAAAATTGTTAATATACCTCTATACTTTAACGTCAAGGAGAAAAAACTATAATGACTAAATCTCATTCAGAAGAAGTGATTGTACCTGAGTTCAATTCTAGCGCAAAGGAATTACCAAGACCATTGGCCGAAAAGTGCCCGAGCATAATTAAGAAATTTATAAGCGCTTATGATGCTAAACCGGATTTTGTTGCTAGATCGCCTGGTAGAGTCAATCTAATTGGTGAACATATTGATTATTGTGACTTCTCGGTTTTACCTTTAGCTATTGATTTTGATATGCTTTGCGCCGTCAAAGTTTTGAACGATGAGATTTCAAGTCTTAAAGCTATATCAGAGGGCTAAGCATGTGTATTCTGAATCTTTAAGAGTCTTGAAGGCTGTGAAATTAATGACTACAGCGAGCTTTACTGCCGACGAAGACTTTTTCAAGCAATTTGGTGCCTTGATGAACGAGTCTCAAGCTTCTTGCGATAAACTTTACGAATGTTCTTGTCCAGAGATTGACAAAATTTGTTCCATTGCTTTGTCAAATGGATCATATGGTTCCCGTTTGACCGGAGCTGGCTGGGGTGGTTGTACTGTTCACTTGGTTCCAGGGGGCCCAAATGGCAACATAGAAAAGGTAAAAGAAGCCCTTGCCAATGAGTTCTACAAGGTCAAGTACCCTAAGATCACTGATGCTGAGCTAGAAAATGCTATCATCGTCTCTAAACCAGCATTGGGCAGCTGTCTATATGAATTAGTCAAGTATACTTCTTTTTTTTACTTTGTTCAGAACAACTTCTCATTTTTTTCTACTCATAACTTTAGCATCACAAAATACGCAATAATAACGAGTAGTAACACTTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCACAAACTTTAAAACACAGGGACAAAATTCTTGATATGCTTTCAACCGCTGCGTTTTGGATACCTATTCTTGACATGATATGACTACCATTTTGTTATTGTACGTGGGGCAGTTGACGTCTTATCATATGTCAAAG...TTGCGAAGTTCTTGGCAAGTTGCCAACTGACGAGATGCAGTAACACTTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCACAAACTTTAAAACACAGGGACAAAATTCTTGATATGCTTTCAACCGCTGCGTTTTGGATACCTATTCTTGACATGATATGACTACCATTTTGTTATTGTACGTGGGGCAGTTGACGTCTTATCATATGTCAAAGTCATTTGCGAAGTTCTTGGCAAGTTGCCAACTGACGAGATGCAGTTTCCTACGCATAATAAGAATAGGAGGGAATATCAAGCCAGACAATCTATCATTACATTTAAGCGGCTCTTCAAAAAGATTGAACTCTCGCCAACTTATGGAATCTTCCAATGAGACCTTTGCGCCAAATAATGTGGATTTGGAAAAAGAGTATAAGTCATCTCAGAGTAATATAACTACCGAAGTTTATGAGGCATCGAGCTTTGAAGAAAAAGTAAGCTCAGAAAAACCTCAATACAGCTCATTCTGGAAGAAAATCTATTATGAATATGTGGTCGTTGACAAATCAATCTTGGGTGTTTCTATTCTGGATTCATTTATGTACAACCAGGACTTGAAGCCCGTCGAAAAAGAAAGGCGGGTTTGGTCCTGGTACAATTATTGTTACTTCTGGCTTGCTGAATGTTTCAATATCAACACTTGGCAAATTGCAGCTACAGGTCTACAACTGGGTCTAAATTGGTGGCAGTGTTGGATAACAATTTGGATTGGGTACGGTTTCGTTGGTGCTTTTGTTGTTTTGGCCTCTAGAGTTGGATCTGCTTATCATTTGTCATTCCCTATATCATCTAGAGCATCATTCGGTATTTTCTTCTCTTTATGGCCCGTTATTAACAGAGTCGTCATGGCCATCGTTTGGTATAGTGTCCAAGCTTATATTGCGGCAACTCCCGTATCATTAATGCTGAAATCTATCTTTGGAAAAGATTTACAATGATTGTACGTGGGGCAGTTGACGTCTTATCATATGTCAAAGTCATTTGCGAAGTTCTTGGCAAGTTGCCAACTGACGAGATGCAGTAACACTTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCACAAACTTTAAAACACAGGGACAAAATTCTTGATATGCTTTCAACCGCTGCGTTTTGGATACCTATTCTTGACATGATATGACTACCATTTTGTTATTGTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATTTATATTGAATTTTCAAAAATTCTTACTTTTTTTTTGGATGGACGCAAAGAAGTTTAATAATCATATTACATGGCATTACCACCATATACATATCCATATCTAATCTTACTTATATGTTGTGGAAATGTAAAGAGCCCCATTATCTTAGCCTAAAAAAACCTTCTCTTTGGAACTTTCAGTAATACGCTTAACTGCTCATTGCTATATTGAAGTACGGATTAGAAGCCGCCGAGCGGGCGACAGCCCTCCGACGGAAGACTCTCCTCCGTGCGTCCTCGTCTTCACCGGTCGCGTTCCTGAAACGCAGATGTGCCTCGCGCCGCACTGCTCCGAACAATAAAGATTCTACAATACTAGCTTTTATGGTTATGAAGAGGAAAAATTGGCAGTAACCTGGCCCCACAAACCTTCAAATTAACGAATCAAATTAACAACCATAGGATGATAATGCGATTAGTTTTTTAGCCTTATTTCTGGGGTAATTAATCAGCGAAGCGATGATTTTTGATCTATTAACAGATATATAAATGGAAAAGCTGCATAACCACTTTAACTAATACTTTCAACATTTTCAGTTTGTATTACTTCTTATTCAAATGTCATAAAAGTATCAACAAAAAATTGTTAATATACCTCTATACTTTAACGTCAAGGAGAAAAAACTATAATGACTAAATCTCATTCAGAAGAAGTGATTGTACCTGAGTTCAATTCTAGCGCAAAGGAATTACCAAGACCATTGGCCGAAAAGTGCCCGAGCATAATTAAGAAATTTATAAGCGCTTATGATGCTAAACCGGATTTTGTTGCTAGATCGCCTGGTAGAGTCAATCTAATTGGTGAACATATTGATTATTGTGACTTCTCGGTTTTACCTTTAGCTATTGATTTTGATATGCTTTGCGCCGTCAAAGTTTTGAACGATGAGATTTCAAGTCTTAAAGCTATATCAGAGGGCTAAGCATGTGTATTCTGAATCTTTAAGAGTCTTGAAGGCTGTGAAATTAATGACTACAGCGAGCTTTACTGCCGACGAAGACTTTTTCAAGCAATTTGGTGCCTTGATGAACGAGTCTCAAGCTTCTTGCGATAAACTTTACGAATGTTCTTGTCCAGAGATTGACAAAATTTGTTCCATTGCTTTGTCAAATGGATCATATGGTTCCCGTTTGACCGGAGCTGGCTGGGGTGGTTGTACTGTTCACTTGGTTCCAGGGGGCCCAAATGGCAACATAGAAAAGGTAAAAGAAGCCCTTGCCAATGAGTTCTACAAGGTCAAGTACCCTAAGATCACTGATGCTGAGCTAGAAAATGCTATCATCGTCTCTAAACCAGCATTGGGCAGCTGTCTATATGAATTAGTCAAGTATACTTCTTTTTTTTACTTTGTTCAGAACAACTTCTCATTTTTTTCTACTCATAACTTTAGCATCACAAAATACGCAATAATAACGAGTAGTAACACTTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCACAAACTTTAAAACACAGGGACAAAATTCTTGATATGCTTTCAACCGCTGCGTTTTGGATACCTATTCTTGACATGATATGACTACCATTTTGTTATTGTACGTGGGGCAGTTGACGTCTTATCATATGTCAAAG...TTGCGAAGTTCTTGGCAAGTTGCCAACTGACGAGATGCAGTAACACTTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCACAAACTTTAAAACACAGGGACAAAATTCTTGATATGCTTTCAACCGCTGCGTTTTGGATACCTATTCTTGACATGATATGACTACCATTTTGTTATTGTACGTGGGGCAGTTGACGTCTTATCATATGTCAAAGTCATTTGCGAAGTTCTTGGCAAGTTGCCAACTGACGAGATGCAGTTTCCTACGCATAATAAGAATAGGAGGGAATATCAAGCCAGACAATCTATCATTACATTTAAGCGGCTCTTCAAAAAGATTGAACTCTCGCCAACTTATGGAATCTTCCAATGAGACCTTTGCGCCAAATAATGTGGATTTGGAAAAAGAGTATAAGTCATCTCAGAGTAATATAACTACCGAAGTTTATGAGGCATCGAGCTTTGAAGAAAAAGTAAGCTCAGAAAAACCTCAATACAGCTCATTCTGGAAGAAAATCTATTATGAATATGTGGTCGTTGACAAATCAATCTTGGGTGTTTCTATTCTGGATTCATTTATGTACAACCAGGACTTGAAGCCCGTCGAAAAAGAAAGGCGGGTTTGGTCCTGGTACAATTATTGTTACTTCTGGCTTGCTGAATGTTTCAATATCAACACTTGGCAAATTGCAGCTACAGGTCTACAACTGGGTCTAAATTGGTGGCAGTGTTGGATAACAATTTGGATTGGGTACGGTTTCGTTGGTGCTTTTGTTGTTTTGGCCTCTAGAGTTGGATCTGCTTATCATTTGTCATTCCCTATATCATCTAGAGCATCATTCGGTATTTTCTTCTCTTTATGGCCCGTTATTAACAGAGTCGTCATGGCCATCGTTTGGTATAGTGTCCAAGCTTATATTGCGGCAACTCCCGTATCATTAATGCTGAAATCTATCTTTGGAAAAGATTTACAATGATTGTACGTGGGGCAGTTGACGTCTTATCATATGTCAAAGTCATTTGCGAAGTTCTTGGCAAGTTGCCAACTGACGAGATGCAGTAACACTTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCACAAACTTTAAAACACAGGGACAAAATTCTTGATATGCTTTCAACCGCTGCGTTTTGGATACCTATTCTTGACATGATATGACTACCATTTTGTTATTGTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAAT

4. TTATATTGAATTTTCAAAAATTCTTACTTTTTTTTTGGATGGACGCAAAGAAGTTTAATAATCATATTACATGGCATTACCACCATATACATATCCATATCTAATCTTACTTATATGTTGTGGAAATGTAAAGAGCCCCATTATCTTAGCCTAAAAAAACCTTCTCTTTGGAACTTTCAGTAATACGCTTAACTGCTCATTGCTATATTGAAGTACGGATTAGAAGCCGCCGAGCGGGCGACAGCCCTCCGACGGAAGACTCTCCTCCGTGCGTCCTCGTCTTCACCGGTCGCGTTCCTGAAACGCAGATGTGCCTCGCGCCGCACTGCTCCGAACAATAAAGATTCTACAATACTAGCTTTTATGGTTATGAAGAGGAAAAATTGGCAGTAACCTGGCCCCACAAACCTTCAAATTAACGAATCAAATTAACAACCATAGGATGATAATGCGATTAGTTTTTTAGCCTTATTTCTGGGGTAATTAATCAGCGAAGCGATGATTTTTGATCTATTAACAGATATATAAATGGAAAAGCTGCATAACCACTTTAACTAATACTTTCAACATTTTCAGTTTGTATTACTTCTTATTCAAATGTCATAAAAGTATCAACAAAAAATTGTTAATATACCTCTATACTTTAACGTCAAGGAGAAAAAACTATAATGACTAAATCTCATTCAGAAGAAGTGATTGTACCTGAGTTCAATTCTAGCGCAAAGGAATTACCAAGACCATTGGCCGAAAAGTGCCCGAGCATAATTAAGAAATTTATAAGCGCTTATGATGCTAAACCGGATTTTGTTGCTAGATCGCCTGGTAGAGTCAATCTAATTGGTGAACATATTGATTATTGTGACTTCTCGGTTTTACCTTTAGCTATTGATTTTGATATGCTTTGCGCCGTCAAAGTTTTGAACGATGAGATTTCAAGTCTTAAAGCTATATCAGAGGGCTAAGCATGTGTATTCTGAATCTTTAAGAGTCTTGAAGGCTGTGAAATTAATGACTACAGCGAGCTTTACTGCCGACGAAGACTTTTTCAAGCAATTTGGTGCCTTGATGAACGAGTCTCAAGCTTCTTGCGATAAACTTTACGAATGTTCTTGTCCAGAGATTGACAAAATTTGTTCCATTGCTTTGTCAAATGGATCATATGGTTCCCGTTTGACCGGAGCTGGCTGGGGTGGTTGTACTGTTCACTTGGTTCCAGGGGGCCCAAATGGCAACATAGAAAAGGTAAAAGAAGCCCTTGCCAATGAGTTCTACAAGGTCAAGTACCCTAAGATCACTGATGCTGAGCTAGAAAATGCTATCATCGTCTCTAAACCAGCATTGGGCAGCTGTCTATATGAATTAGTCAAGTATACTTCTTTTTTTTACTTTGTTCAGAACAACTTCTCATTTTTTTCTACTCATAACTTTAGCATCACAAAATACGCAATAATAACGAGTAGTAACACTTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCACAAACTTTAAAACACAGGGACAAAATTCTTGATATGCTTTCAACCGCTGCGTTTTGGATACCTATTCTTGACATGATATGACTACCATTTTGTTATTGTACGTGGGGCAGTTGACGTCTTATCATATGTCAAAG...TTGCGAAGTTCTTGGCAAGTTGCCAACTGACGAGATGCAGTAACACTTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCACAAACTTTAAAACACAGGGACAAAATTCTTGATATGCTTTCAACCGCTGCGTTTTGGATACCTATTCTTGACATGATATGACTACCATTTTGTTATTGTACGTGGGGCAGTTGACGTCTTATCATATGTCAAAGTCATTTGCGAAGTTCTTGGCAAGTTGCCAACTGACGAGATGCAGTTTCCTACGCATAATAAGAATAGGAGGGAATATCAAGCCAGACAATCTATCATTACATTTAAGCGGCTCTTCAAAAAGATTGAACTCTCGCCAACTTATGGAATCTTCCAATGAGACCTTTGCGCCAAATAATGTGGATTTGGAAAAAGAGTATAAGTCATCTCAGAGTAATATAACTACCGAAGTTTATGAGGCATCGAGCTTTGAAGAAAAAGTAAGCTCAGAAAAACCTCAATACAGCTCATTCTGGAAGAAAATCTATTATGAATATGTGGTCGTTGACAAATCAATCTTGGGTGTTTCTATTCTGGATTCATTTATGTACAACCAGGACTTGAAGCCCGTCGAAAAAGAAAGGCGGGTTTGGTCCTGGTACAATTATTGTTACTTCTGGCTTGCTGAATGTTTCAATATCAACACTTGGCAAATTGCAGCTACAGGTCTACAACTGGGTCTAAATTGGTGGCAGTGTTGGATAACAATTTGGATTGGGTACGGTTTCGTTGGTGCTTTTGTTGTTTTGGCCTCTAGAGTTGGATCTGCTTATCATTTGTCATTCCCTATATCATCTAGAGCATCATTCGGTATTTTCTTCTCTTTATGGCCCGTTATTAACAGAGTCGTCATGGCCATCGTTTGGTATAGTGTCCAAGCTTATATTGCGGCAACTCCCGTATCATTAATGCTGAAATCTATCTTTGGAAAAGATTTACAATGATTGTACGTGGGGCAGTTGACGTCTTATCATATGTCAAAGTCATTTGCGAAGTTCTTGGCAAGTTGCCAACTGACGAGATGCAGTAACACTTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCACAAACTTTAAAACACAGGGACAAAATTCTTGATATGCTTTCAACCGCTGCGTTTTGGATACCTATTCTTGACATGATATGACTACCATTTTGTTATTGTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATTTTTATATTGAATTTTCAAAAATTCTTACTTTTTTTTTGGATGGACGCAAAGAAGTTTAATAATCATATTACATGGCATTACCACCATATACATATCCATATCTAATCTTACTTATATGTTGTGGAAATGTAAAGAGCCCCATTATCTTAGCCTAAAAAAACCTTCTCTTTGGAACTTTCAGTAATACGCTTAACTGCTCATTGCTATATTGAAGTACGGATTAGAAGCCGCCGAGCGGGCGACAGCCCTCCGACGGAAGACTCTCCTCCGTGCGTCCTCGTCTTCACCGGTCGCGTTCCTGAAACGCAGATGTGCCTCGCGCCGCACTGCTCCGAACAATAAAGATTCTACAATACTAGCTTTTATGGTTATGAAGAGGAAAAATTGGCAGTAACCTGGCCCCACAAACCTTCAAATTAACGAATCAAATTAACAACCATAGGATGATAATGCGATTAGTTTTTTAGCCTTATTTCTGGGGTAATTAATCAGCGAAGCGATGATTTTTGATCTATTAACAGATATATAAATGGAAAAGCTGCATAACCACTTTAACTAATACTTTCAACATTTTCAGTTTGTATTACTTCTTATTCAAATGTCATAAAAGTATCAACAAAAAATTGTTAATATACCTCTATACTTTAACGTCAAGGAGAAAAAACTATAATGACTAAATCTCATTCAGAAGAAGTGATTGTACCTGAGTTCAATTCTAGCGCAAAGGAATTACCAAGACCATTGGCCGAAAAGTGCCCGAGCATAATTAAGAAATTTATAAGCGCTTATGATGCTAAACCGGATTTTGTTGCTAGATCGCCTGGTAGAGTCAATCTAATTGGTGAACATATTGATTATTGTGACTTCTCGGTTTTACCTTTAGCTATTGATTTTGATATGCTTTGCGCCGTCAAAGTTTTGAACGATGAGATTTCAAGTCTTAAAGCTATATCAGAGGGCTAAGCATGTGTATTCTGAATCTTTAAGAGTCTTGAAGGCTGTGAAATTAATGACTACAGCGAGCTTTACTGCCGACGAAGACTTTTTCAAGCAATTTGGTGCCTTGATGAACGAGTCTCAAGCTTCTTGCGATAAACTTTACGAATGTTCTTGTCCAGAGATTGACAAAATTTGTTCCATTGCTTTGTCAAATGGATCATATGGTTCCCGTTTGACCGGAGCTGGCTGGGGTGGTTGTACTGTTCACTTGGTTCCAGGGGGCCCAAATGGCAACATAGAAAAGGTAAAAGAAGCCCTTGCCAATGAGTTCTACAAGGTCAAGTACCCTAAGATCACTGATGCTGAGCTAGAAAATGCTATCATCGTCTCTAAACCAGCATTGGGCAGCTGTCTATATGAATTAGTCAAGTATACTTCTTTTTTTTACTTTGTTCAGAACAACTTCTCATTTTTTTCTACTCATAACTTTAGCATCACAAAATACGCAATAATAACGAGTAGTAACACTTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCACAAACTTTAAAACACAGGGACAAAATTCTTGATATGCTTTCAACCGCTGCGTTTTGGATACCTATTCTTGACATGATATGACTACCATTTTGTTATTGTACGTGGGGCAGTTGACGTCTTATCATATGTCAAAG...TTGCGAAGTTCTTGGCAAGTTGCCAACTGACGAGATGCAGTAACACTTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCACAAACTTTAAAACACAGGGACAAAATTCTTGATATGCTTTCAACCGCTGCGTTTTGGATACCTATTCTTGACATGATATGACTACCATTTTGTTATTGTACGTGGGGCAGTTGACGTCTTATCATATGTCAAAGTCATTTGCGAAGTTCTTGGCAAGTTGCCAACTGACGAGATGCAGTTTCCTACGCATAATAAGAATAGGAGGGAATATCAAGCCAGACAATCTATCATTACATTTAAGCGGCTCTTCAAAAAGATTGAACTCTCGCCAACTTATGGAATCTTCCAATGAGACCTTTGCGCCAAATAATGTGGATTTGGAAAAAGAGTATAAGTCATCTCAGAGTAATATAACTACCGAAGTTTATGAGGCATCGAGCTTTGAAGAAAAAGTAAGCTCAGAAAAACCTCAATACAGCTCATTCTGGAAGAAAATCTATTATGAATATGTGGTCGTTGACAAATCAATCTTGGGTGTTTCTATTCTGGATTCATTTATGTACAACCAGGACTTGAAGCCCGTCGAAAAAGAAAGGCGGGTTTGGTCCTGGTACAATTATTGTTACTTCTGGCTTGCTGAATGTTTCAATATCAACACTTGGCAAATTGCAGCTACAGGTCTACAACTGGGTCTAAATTGGTGGCAGTGTTGGATAACAATTTGGATTGGGTACGGTTTCGTTGGTGCTTTTGTTGTTTTGGCCTCTAGAGTTGGATCTGCTTATCATTTGTCATTCCCTATATCATCTAGAGCATCATTCGGTATTTTCTTCTCTTTATGGCCCGTTATTAACAGAGTCGTCATGGCCATCGTTTGGTATAGTGTCCAAGCTTATATTGCGGCAACTCCCGTATCATTAATGCTGAAATCTATCTTTGGAAAAGATTTACAATGATTGTACGTGGGGCAGTTGACGTCTTATCATATGTCAAAGTCATTTGCGAAGTTCTTGGCAAGTTGCCAACTGACGAGATGCAGTAACACTTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCACAAACTTTAAAACACAGGGACAAAATTCTTGATATGCTTTCAACCGCTGCGTTTTGGATACCTATTCTTGACATGATATGACTACCATTTTGTTATTGTTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATAATGTTTTCAATGTAAGAGATTTCGATTATCTTATAGTTCATACATGCTTCAACTACTTAATAAATGATTGTATGATTT Exons Promoter motifs 3’ UTR motifs Introns

5. Comparing genomes reveals functional elements • Protein-coding genes • Ultra-conserved elements • Short regulatory motifs

6. Regulatory Motif Discovery • Gene regulation • Genes are turned on / off in response to changing environments • Gene regulatory logic is controlled by sequence motifs • Specialized proteins (transcription factors) recognize motifs • What makes motif discovery hard? • Motifs are short (6-8 bp) and usually degenerate • Act at variable distances upstream (or downstream) of target gene GAL1 Gal4 Gal4 Mig1 ATGACTAAATCTCATTCAGAAGAAGTGA CGG CCG CGG CCG CCCCW

7. Regulatory Motif Discovery Study known motifs Derive conservation rules Discover novel motifs

8. Known motifs are preferentially conserved Is this enough to discover motifs? Is this enough to discover motifs? No.

9. human CTCTTAATGGTACACGTTCTGCCT----AAGTAGCCTAGACGCTCCCGTGCGCCC-GGGGdog CTCTTA-CGGGGCACATTCTGCTTTCAACAGTGGGGCAGACGGTCCCGCGCGCCCCAAGGmouse GTCTTAGGAGGCT-CGATCGCC---------------------GCCTGCATTATT-----rat GTCTTAGTTGGCCACGACCTGC---------------------TCATGCATAATT----- ***** * * * * * * human CGGGTAGGCCTGGCCGAAAATCTCTCCCGCGCGCCTGACCTTGGGTTGCCCCAGCCAGGCdog CAGGC---CCGGGCTGCAGACCTGCCCTGAGGGAATGACCTTGGGCGGCCGCAGCGGGGCmouse --------------CACAAGCCTGTGGCGCGC-CGTGACCTTGGGCTGCCCCAGGCGGGCrat --------------CACAAGTTTCTC---TGC-CCTGACCTTGGGTTGCCCCAGGCGAG- * * * ********** *** *** * human TGCGGGCCCGAGACCCCCG-------------------GGCCTCCCTGCCCCCCGCGCCGdog CGCGGGCCCAGGCCCCCCTCCCTCCCTCCCTCCCTCCCTCCCTCCCTGCCCCCCGGACCGmouse TGCAGGCTCACCACCCCGTCTTTTCT---------------------GCTTTTCGAGTCGrat -GCATACACCCCGCCTTTTTTTTTTTTTT---------TTTTTTTTTGCCGTTCAAG-AG ** * * ** ** * * Erra Known motifs are preferentially conserved human CTCTTAATGGTACACGTTCTGCCT----AAGTAGCCTAGACGCTCCCGTGCGCCC-GGGGdog CTCTTA-CGGGGCACATTCTGCTTTCAACAGTGGGGCAGACGGTCCCGCGCGCCCCAAGGmouse GTCTTAGGAGGCT-CGATCGCC---------------------GCCTGCATTATT-----rat GTCTTAGTTGGCCACGACCTGC---------------------TCATGCATAATT----- ***** * * * * * * human CGGGTAGGCCTGGCCGAAAATCTCTCCCGCGCGCCTGACCTTGGGTTGCCCCAGCCAGGCdog CAGGC---CCGGGCTGCAGACCTGCCCTGAGGGAATGACCTTGGGCGGCCGCAGCGGGGCmouse --------------CACAAGCCTGTGGCGCGC-CGTGACCTTGGGCTGCCCCAGGCGGGCrat --------------CACAAGTTTCTC---TGC-CCTGACCTTGGGTTGCCCCAGGCGAG- * * * ********** *** *** * human TGCGGGCCCGAGACCCCCG-------------------GGCCTCCCTGCCCCCCGCGCCGdog CGCGGGCCCAGGCCCCCCTCCCTCCCTCCCTCCCTCCCTCCCTCCCTGCCCCCCGGACCGmouse TGCAGGCTCACCACCCCGTCTTTTCT---------------------GCTTTTCGAGTCGrat -GCATACACCCCGCCTTTTTTTTTTTTTT---------TTTTTTTTTGCCGTTCAAG-AG ** * * ** ** * * human CTCTTAATGGTACACGTTCTGCCT----AAGTAGCCTAGACGCTCCCGTGCGCCC-GGGGdog CTCTTA-CGGGGCACATTCTGCTTTCAACAGTGGGGCAGACGGTCCCGCGCGCCCCAAGGmouse GTCTTAGGAGGCT-CGATCGCC---------------------GCCTGCATTATT-----rat GTCTTAGTTGGCCACGACCTGC---------------------TCATGCATAATT----- ***** * * * * * * human CGGGTAGGCCTGGCCGAAAATCTCTCCCGCGCGCCTGACCTTGGGTTGCCCCAGCCAGGCdog CAGGC---CCGGGCTGCAGACCTGCCCTGAGGGAATGACCTTGGGCGGCCGCAGCGGGGCmouse --------------CACAAGCCTGTGGCGCGC-CGTGACCTTGGGCTGCCCCAGGCGGGCrat --------------CACAAGTTTCTC---TGC-CCTGACCTTGGGTTGCCCCAGGCGAG- * * * ********** *** *** * human TGCGGGCCCGAGACCCCCG-------------------GGCCTCCCTGCCCCCCGCGCCGdog CGCGGGCCCAGGCCCCCCTCCCTCCCTCCCTCCCTCCCTCCCTCCCTGCCCCCCGGACCGmouse TGCAGGCTCACCACCCCGTCTTTTCT---------------------GCTTTTCGAGTCGrat -GCATACACCCCGCCTTTTTTTTTTTTTT---------TTTTTTTTTGCCGTTCAAG-AG ** * * ** ** * * Gabpa Is this enough to discover motifs? No

10. Erra Erra Erra Dog Mouse Rat Conservation rate: 37% Known motifs are frequently conserved Human • Across the human promoter regions, the Erra motif: • appears 434 times • is conserved 162 times • Compare to random control motifs • Conservation rate of control motifs: 6.8% • Erra enrichment: 5.4-fold • Erra p-value < 10-50 (25 standard deviations under binomial) Motif Conservation Score (MCS)

11. MCS distribution of all 6-mers shows excess conservation High scoring patterns include known motifs Excess specific to promoters and 3’-UTRs (not introns) For MCS > 6, estimate 97% specificity Select motifs with MCS > 6.0, cluster Motif density Motif density Motif Conservation Score (MCS)

12. Hill-climbing in sequence space • Seed selection • Three mini-motif conservation criteria (CC1, CC2, CC3) • Motif extension • Non-random conservation of neighbors • Motif collapsing • Merge neighbors using hierarchical clustering, avg-max-linkage • Re-scoring complex motifs • Motif conservation score for full motifs (MCS)

13. CGG-11-CCG Test 1: Intergenic conservation Conserved count Total count

14. N r Conservation rate Binomial score Test 1: Selecting mini-motifs • Estimate basal rate of conservation • Expected conservation rate at the evolutionary distances observed • Average conservation rate of non-outlier mini-motifs • Score conservation of mini-motif • k: conserved motif occurrences • n: total motif occurrences • r: basal conservation rate • Evaluate binomial probability of observing k successes out of n trials • Assign z-score to each mini-motif • Bulk of distribution is symmetric • Estimate specificity as (R-L)/R • Select cutoff: 5.0 sigma • 1190 mini-motifs, 97.5% non-random Specificity Cutoff Right tail Left tail

15. CGG-11-CCG Higher Conservation in Genes Test 2: Intergenic vs. Coding Intergenic Conservation Coding Conservation

16. CGG-11-CCG Downstream motifs? Most Patterns Test 3: Upstream vs. Downstream Upstream Conservation Downstream Conservation

17. 5 6 Y R T C G C A C G A Extend Extend Extend Collapse Extend G T C A C A C G A A T C R Y A C G A Collapse Collapse Collapse R T C G C A C G A Merge 72 Full motifs Full Motifs Constructing full motifs Test 1 Test 2 Test 3 2,000 Mini-motifs R T C A A C G R

18. Find maximally discriminating neighborhood N1 M1 R T C A G A C G W N2 M2 Y T C x H A x G S • Evaluate non-randomness of neighborhood • chi-square contingency test on [N1,M1], [N2,M2] Extending mini-motifs • Separate conserved and non-conserved instances 6 T C A A C G Causal set 6 T C x A x G Random set

19. 174 motifs in promoters 106 motifs in 3’ UTRs Systematically test candidate patterns gap S R T G C Y W T A G R • Enumerate • Length between 6 and 15 nt, allow central gap • 11 letter alphabet (A C G T, 2-fold codes, N) • Score • Compute binomial score (conserved vs. total) • Select MCS > 6.0  specificity 97% • Cluster • Sequence similarity • Overlapping occurrences All potential motifs Evaluate MCS Cluster similar motifs Are these real ?

20. Functions of discovered motifs

21. Evidence of motif function Promoter 3’-UTR • Promoter motifs: • Comparison to known motifs • Distance from TSS • Expression enrichment Stop ATG 174 motifs 106 motifs

22. Promoter motifs match known TF binding sites Compare discovered motifs to TRANSFAC database of 125 known motifs 45% of discovered motifs match TRANSFAC motifs (only 2% of control sequences match TRANSFAC motifs) 55% of TRANSFAC motifs match discovered motifs

23. (2) Promoter motifs show preferred distance to TSS Motif instances in human Conserved motif sites in all four species Motif 4 -81 Each of 174 discovered motifs Motif 8 -63 Distance from TSS Discovered motifs occur preferentially Within 200 bp of Transcription Start Site Individual motifs show strong peaks Regardless of conservation 32% of discovered motifs show strong positional bias

24. (3) Promoter motifs enriched in specific tissues New motifs Known TFs 70% of motifs show significant enrichment in at least one tissue

25. New New New New New Summary for promoter motifs • 174 promoter motifs • 70 match known TF motifs • 115 expression enrichment • 60 show positional bias  75% have evidence • Control sequences < 2% match known TF motifs < 5% expression enrichment < 3% show positional bias  < 7% false positives Most discovered motifs are likely to be functional

26. Summary of Promoter Motifs

27. Similar analysis in 5% most conserved regions in human 12-22 bp long motifs

28. Similar analysis in 5% most conserved regions in human

29. Overview of Motif Discovery Algorithms

30. Motif Representation GTATAA CTATAA GTCTTA ATATAC GTAATA TTGTAC GTATTA GTATTC ATCTAA GTATAM IUPAC Complex Dependency Graphical Models ATATAC GTAATA ATCTAA GTATTC GTATAA GTATAA CTATAA GTATTA Consensus PSSM TTGTAC GTCTTA Nonparametric – Graph or Bag of Words

31. Motif Representation – Pairwise Dependencies Complex Dependency Graphical Models

32. Motif Representation – MotifScan ATATAC GTAATA ATCTAA GTATTC GTATAA CTATAA GTATTA TTGTAC GTCTTA

33. Motif Finding • Given a set of promoter sequences • For example, common expression pattern of the respective genes in microarrays ACCGAGAGTATAAGCTTACGTGACTTGCATGATCTTGCGATGTGTGTTCAGCT ATCGTACGTTGAGGAGAGGCGGTAATAGAAGTACGTCGATGTCGTCGTACAT TTCCTATAAGATCGACTGTAGGGAGAGTCTCTGAGAGTATTGCTGGCATGTG ACTTCGAGGAGAGATTCTCTAGATCTATGCTGTGGTATTAAGAGATCTCTAG ATCGATGCGCTGATCGCTATAATATATCGGCGGTATCTGGTTGATCTGGTGT GACTGATGTATCGTATCTGATCTGTCGGTATAATATAGCTGTCTGATTAGTTG TCTCTAGATGCTGTGCTGATGGTCTTATCGATGTGCGACGGTAATAGTATCCT • Find a common motif that they share GTATAA GTAATA CTATAA GTATTA CTATAA GTATAA GTAATA

34. Most Popular Approaches • Expectation Maximization – MEME • Sequences are mixtures of • Motif model M, e.g., a motif PSSM • Background model B, e.g., 3rd order model of promoters • Learn model by • Starting from random M, learned B from promoters • Assign each position in input to M or B, accordingly • Re-estimate M and B based on current assignments • Gibbs Sampling – AlignACE, BioProspector • Update 1-seq x at a time • Remove from M • Pick a new location in x based on M x M

35. MotifCut • Construct a graph of all promoters • Each k-mer in each promoter is a node • Nodes are connected with edges of weight proportional to sequence similarity • Find maximum density subgraph … ACAGGAT CAGGATC AGGATCA GGATCAC • ACAGGATCACTGATGCAGCATGCATGCATCG • CTAGTCGTAGTCTCGATCTAGCTGTGTGTC • CATGATGCGCGATCTTGCTGTGGTCATTAGC • ATCGAGGCGAGAGAGATCTCTCTAGTGTACT