Jonathan Eisen talk on "Phylogenomics of Microbes" at Lake Arrowhead Small Genomes Meeting 2002
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Talk by Jonathan Eisen on Phylogenomics of microbes at Lake Arrowhead Small Genomes meeting in 2002.
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Jonathan Eisen talk on "Phylogenomics of Microbes" at Lake Arrowhead Small Genomes Meeting 2002
- 1. TIGR
- 2. “Nothing in biology makes sense except in the light of evolution.” T. H. Dobzhansky (1973) TIGR
- 3. Topics of Discussion • Introduction to phylogenomics • Uses of evolutionary analysis in genomics – Selection of species – Functional prediction – Gene duplication – Gene loss – Genome rearrangements – Lateral transfer – Uncultured species – Specialization TIGR
- 4. Phylogenomic Analysis Phylogenomics involves combining evolutionary reconstructions of genes, proteins, pathways, and species with analysis of complete genome sequences. TIGR
- 5. Uses of Phylogenomics • Selection of species • Functional prediction • Gene duplication • Intragenomic movement • Gene loss • Lateral transfer • Genome rearrangements • Uncultured species TIGR
- 6. Strain Selection and Evolution • Increasing phylogenetic representation • Determining relatedness to model organism • Understanding major evolutionary transitions • Identifying taxa with unusual (high or low) rates of evolution • Identifying source of DNA from uncultured species • Species naming and type strains (e.g., see Ward et. al. 2001) TIGR
- 7. Evolutionary Diversity Still Poorly Represented in Complete Genomes TIGR Bacteria Archaea
- 8. S. pombe Genome Analysis Eukaryotes vs. Prokaryotes S. pombe S. cerevisiae Eukaryotes Encephalatozoon Archaea Worm Fly Bacteria Humans Dictyostelium Arabidopsis Chlamydomonas Phytophthora Tetrahymena Plasmodium Trypanosoma Euglena Naegleria Trichomonas Giardia TIGR
- 9. Single vs. Multi-celled S. pombe Fungi S. cerevisiae Encephalatozoon Microsporidia Worm Animals Fly Humans Dictyostelium Dictyostelia Arabidopsis Plants Chlamydomonas Phytophthora Heterokonts Tetrahymena Ciliates Plasmodium Apicomplexa Trypanosoma Kinetoplastids Euglena Euglenas Naegleria Acrasidae Trichomonas Parabisalia TIGR Giardia Diplomonads
- 10. Uses of Phylogenomics • Selection of species • Functional prediction • Gene duplication • Intragenomic movement • Gene loss • Lateral transfer • Genome rearrangements • Uncultured species TIGR
- 11. Predicting Function • Identification of motifs • Homology/similarity based methods – Highest hit, top hit, HMMs, threading • Evolutionary methods – Phylogenetic trees – Ds/Dn – Phylogenetic profiles TIGR
- 12. FlyMutS.Borbu TrepaGTBP.Mouse hMSH4.Human MSH4.Caeel MSH4.Yeast orf.Arath hMSH3.Human orf.Chltr orf.Deira atMSH2.Arath MSH2.Neucr MSH2.Yeast MSH2.Human MSH2.Mouse MSH2.Rat MSH2.Xenla SPE1.Drome orf.Trepa MutS.Aquae orfStrpy MutS.Helpy yshD MSH3.Yeast MutS sgMutS.Saugl orfGTBP.Human MutS.Bacsu MSH6.Arath MutS2 orf MutS.Metth hMHS5 MSH5 MutS Swi4.Spombe MSH1.Spombe .MutS Chltr Thema Neigo Arath Neucr Xenla Trepa Theaq Ecoli Bacsu Strpy Yeast Human MouseMSH1.Yeast MSH6.Yeast Rep3.Mouse MutS2.Saugl Helpy Deira Synsp Aquae Borbu MutS2.Metth MutS2-Saugl MutS2-Metth FlyMSH5 Caeel Helpy MutS2.Saugl MutS2.Metth Chltr Deira Theaq Thema Neigo EcoliMSH3 Synsp Bacsu Strpy Borbu ArathMSH6 Neucr Yeast Human Mouse RatMutS1 Xenla Aquae MSH1 Spombe Yeast Mouse Caeel Human Spombe Spombe Yeast Mouse Yeast Human Arath Caeel Spombe Arath MutS2 MSH6 MSH3 MSH2 MSH4 MutS1 MSH1 MSH5 MutS2 MSH2 MSH4 D. C. B. A. Neigo Bacsu Synsp Borbu Deira Strpy Ecoli Aquae Theaq Thema human Yeast Caeel SegAllMMR && Segregationin All MMR regation MMR of Mismatches and in Nucleus Crossover Large Loops (Bacteria) Crossover Mitochondria Small Loops in Nucleus in Nucleus TIGR
- 13. rRNA and Uncultured Microbes TIGR
- 14. Evolutionary Rate Variation 6 5 4 1 3 2 TIGR
- 15. Uses of Phylogenomics • Selection of species • Functional prediction • Gene duplication • Gene loss • Lateral transfer • Genome rearrangements • Uncultured species TIGR
- 16. Why Duplications Are Useful to Identify • Allows division into orthologs and paralogs • Improves functional predictions • Helps identify mechanisms of duplication • Can be used to study mutation processes in different parts of a genome • Lineage specific duplications may be indicative of species’ specific adaptations TIGR
- 17. Lineage Specific Duplications in Wolbachia wMel Annotation hypothetical protein prophage LambdaW2, baseplate ankyrin repeat domain protein hypothetical protein assembly protein J, putative ankyrin repeat domain protein hypothetical protein prophage LambdaW2, baseplate ankyrin repeat domain protein hypothetical protein assembly protein V, putative ankyrin repeat domain protein hypothetical protein FRAMESHIFT ankyrin repeat domain protein hypothetical protein prophage LambdaW2, baseplate ankyrin repeat domain protein hypothetical protein assembly protein V, putative ankyrin repeat domain protein hypothetical protein FRAMESHIFT conserved domain protein hypothetical protein prophage LambdaW2, baseplate conserved domain protein hypothetical protein assembly protein W, putative conserved domain protein hypothetical protein prophage LambdaW2, minor tail conserved domain protein hypothetical protein protein Z, putative, conserved hypothetical protein hypothetical protein FRAMESHIFT conserved hypothetical protein hypothetical protein prophage LambdaW2, site- conserved hypothetical protein hypothetical protein specific recombinase, resolvase conserved hypothetical protein hypothetical protein family conserved hypothetical protein hypothetical protein prophage LambdaW4, ankyrin conserved hypothetical protein hypothetical protein repeat domain protein conserved hypothetical protein hypothetical protein prophage LambdaW4, DNA conserved hypothetical protein hypothetical protein methylase conserved hypothetical protein hypothetical protein prophage LambdaW4, portal conserved hypothetical protein hypothetical protein protein, FRAMESHIFT conserved hypothetical protein hypothetical protein prophage LambdaW4, portal conserved hypothetical protein hypothetical protein protein, FRAMESHIFT conserved hypothetical protein hypothetical protein prophage LambdaW4, terminase conserved hypothetical protein hypothetical protein large subunit, putative conserved hypothetical protein hypothetical protein prophage LambdaW5, ankyrin conserved hypothetical protein hypothetical protein repeat domain protein conserved hypothetical protein hypothetical protein prophage LambdaW5, ankyrin conserved hypothetical protein hypothetical protein repeat domain protein conserved hypothetical protein hypothetical protein prophage LambdaW5, ankyrin conserved hypothetical protein hypothetical protein repeat domain protein conserved hypothetical protein hypothetical protein prophage LambdaW5, baseplate conserved hypothetical protein hypothetical protein assembly protein J, putative, conserved hypothetical protein hypothetical protein FRAMESHIFT conserved hypothetical protein hypothetical protein prophage LambdaW5, baseplate FRAMESHIFT hypothetical protein assembly protein V, putative conserved hypothetical protein hypothetical protein prophage LambdaW5, baseplate POINT MUTATION hypothetical protein assembly protein W, putative conserved hypothetical protein, hypothetical protein prophage LambdaW5, minor tail degenerate hypothetical protein protein Z, putative, degenerate, conserved hypothetical protein, hypothetical protein FRAMESHIFT FRAMESHIFT hypothetical protein prophage LambdaW5, site- conserved hypothetical protein, hypothetical protein specific recombinase, resolvase FRAMESHIFT hypothetical protein family conserved hypothetical protein, hypothetical protein regulatory protein RepA, putative FRAMESHIFT hypothetical protein regulatory protein RepA, putative conserved hypothetical protein, hypothetical protein reverse transcriptase, putative FRAMESHIFT hypothetical protein reverse transcriptase, putative conserved hypothetical protein, hypothetical protein reverse transcriptase, putative interruption-C hypothetical protein sodium/alanine symporter family conserved hypothetical protein, hypothetical protein protein POINT MUTATION hypothetical protein sodium/alanine symporter family conserved hypothetical protein, hypothetical protein protein POINT MUTATION hypothetical protein TenA/THI-4 family protein conserved hypothetical protein, hypothetical protein transcriptional regulator truncated hypothetical protein transcriptional regulator conserved hypothetical protein, hypothetical protein transcriptional regulator truncation hypothetical protein transcriptional regulator DNA mismatch repair protein hypothetical protein transcriptional regulator MutL (mutL) hypothetical protein transcriptional regulator DNA repair protein RadC, hypothetical protein transcriptional regulator, putative putative hypothetical protein translation elongation factor Tu DNA repair protein RadC, hypothetical protein (tuf) putative, truncation hypothetical protein translation elongation factor Tu DNA repair protein RadC, hypothetical protein (tuf) truncation hypothetical protein transposase, degenerate DnaJ domain protein hypothetical protein transposase, IS4 family DnaJ domain protein hypothetical protein transposase, IS4 family exopolysaccharide synthesis hypothetical protein transposase, IS4 family protein ExoD-related protein major facilitator family transposase, IS5 family, exopolysaccharide synthesis transporter interruption-N protein ExoD-related protein major facilitator family transposase, IS5 family, HNH endonuclease family transporter truncation protein major facilitator family transposase, putative, degenerate HNH endonuclease family transporter transposase, putative, degenerate protein membrane protein, putative transposase, putative, degenerate hypothetical protein membrane protein, putative type IV secretion system protein hypothetical protein membrane protein, putative VirB4, putative hypothetical protein MutL family protein UDP-N-acetylglucosamine hypothetical protein Na+/H+ antiporter family protein pyrophosphorylase-related hypothetical protein Na+/H+ antiporter, putative protein hypothetical protein permease, putative hypothetical protein portal protein, FRAMESHIFT hypothetical protein portal protein, FRAMESHIFT hypothetical protein prophage LambdaW1, DNA TIGR hypothetical protein methylase hypothetical protein prophage LambdaW1, terminase hypothetical protein large subunit, putative hypothetical protein prophage LambdaW2, ankyrin hypothetical protein repeat domain protein hypothetical protein prophage LambdaW2, ankyrin hypothetical protein repeat domain protein
- 18. MutL Duplication in Wolbachia wMel ORF01096 DNA mismatch repair protein MutL (mutL) ORF00446 MutL family protein TIGR
- 19. MutL Duplication in Wolbachia wMel TIGR
- 20. Older Duplication of UVDE Schizosaccharomyces pombeGP139 Neurospora crassaPIRS55262S552 Clostridium perfringensGP18145 Bacillus subtilisSPP45864YWJD Bacillus cereusGP6759487embCAB B BACAN 01914 UV endonuclease Bacillus haloduransOMNINTL01BH B BACAN 01459 UV endonuclease Deinococcus radioduransGP61167 Nostoc sp. PCC 7120GP17130610d 0.1 TIGR
- 21. Uses of Phylogenomics • Selection of species • Functional prediction • Gene duplication • Intragenomic movement • Gene loss • Lateral transfer • Genome rearrangements • Uncultured species TIGR
- 22. X-files Eisen et al. 2000. Genome Biology 1(6): 11.1-11.9 Also see Tillier and Collins. 2000. Nature Genetics 26(2):195-7 and Suyama and Bork. 2001. Trends Genetics 17: 10-13. TIGR
- 23. C. trachomatis vs C. pneumoniae Dot Plot Origin C. pneumoniae AR39 Terminus TIGR C. trachomatis MoPn Read et al. 2000
- 24. StrpB vs. StrpA All 13623100 13622900 13622700 13622500 13622300 Series1 13622100 13621900 13621700 13621500 13621300 TIGR 0 500 1000 1500 2000 2500
- 25. StrpB vs. StrpA: Orthologs 13623100 13622900 13622700 13622500 13622300 Series1 13622100 13621900 13621700 13621500 13621300 TIGR 0 500 1000 1500 2000 2500
- 26. Uses of Phylogenomics • Selection of species • Functional prediction • Gene duplication • Intragenomic movement • Gene loss • Lateral transfer • Genome rearrangements • Uncultured species TIGR
- 27. Most ‘Evidence’ for Gene Transfer has Alternative Explanations Observation Other Causes Always Occurs Unusual Distribution Sampling bias Not if recipient already has gene. Unusual GC/Codons Selection Not if donor/recipient similar. Not if it occurred long ago. High hit to "distant" species Selection Usually. Rate variation Gene loss Incongruent trees Bad trees Usually. Missed paralogs Correlation of above with Selection Only if genes keep order after neighbors transfer. TIGR
- 28. Steps in Lateral Gene Transfer A B C D 6 3-5 2 1 TIGR
- 29. Mitochondrial Genome Integration into A. thaliana chrII 0 4E+05 3E+05 2E+05 1E+05 3.6E+06 3.5E+06 3.4E+06 3.3E+06 3.2E+06 thaliana D. C. B. Alternative Chromosome II Possible Mitochondrial Form D’D’A. 1’ 1 A B C A’ 3 Insertion Mitochondrial PointAlternative Genome TIGR Lin et al., 1999
- 30. Number of pBVTs Depends 1800 on # of Genomes Analyzed 1600 1400 1200 Fruit fly 1000 C. elegans Arabidopsis Yeast 800 Parasites 600 400 200 0 1 2 3 4 5 Other TIGR Number of protein sets Salzberg et al. 2001
- 31. Trees Don’t Support Transfer II TIGR
- 32. Uses of Phylogenomics • Selection of species • Functional prediction • Gene duplication • Intragenomic movement • Gene loss • Lateral transfer • Genome rearrangements • Uncultured species TIGR
- 33. TIGR Beja O, et.al., Science 2000 289:1902-6, Nature (2001) 411: 786-789
- 34. Puf Operons from Uncultured Bacteria TIGR
- 35. Puf Operons vs. Cultured Species TIGR
- 36. Alternative Phylogenetic Anchors Chlorobium tepidum Cytophaga hutchinsonii Prevotella ruminocola Bacteroides fragilis Porphyromonas gingivalis MBBAD68TR TIGR MBBAD65TR
- 37. Acknowledgements • Outside TIGR –A. Stoltzfus –H. Ochman –D. Bryant –W. F. Doolittle –M. Eisen –M-I Benito • $$$: –NSF –NIH –ONR –DOE –NEB TIGR
- 38. B. anthracis lineage specific duplications ORF04205 molybdopterin biosynthesis protein MoeA (moeA) ORF05907 molybdopterin biosynthesis protein MoeA (moeA) ORF02636 molybdopterin biosynthesis protein MoeA (moeA) ORF04204 molybdopterin biosynthesis protein MoeB, putative ORF05908 molybdopterin biosynthesis protein MoeB, putative ORF02634 molybdopterin biosynthesis protein MoeB, putative ORF05904 molybdopterin converting factor, subunit 1 (moaD) ORF02639 molybdopterin converting factor, subunit 1 (moaD) ORF04206 molybdopterin converting factor, subunit 2 (moaE) ORF05905 molybdopterin converting factor, subunit 2 (moaE) ORF02638 molybdopterin converting factor, subunit 2 (moaE) TIGR Based on Read et al. submitted
- 39. Schizosaccharomyces pombeGP139 Neurospora crassaPIRS55262S552 Clostridium perfringensGP18145 Bacillus subtilisSPP45864YWJD Bacillus cereusGP6759487embCAB B BACAN 01914 UV endonuclease Bacillus haloduransOMNINTL01BH B BACAN 01459 UV endonuclease Deinococcus radioduransGP61167 Nostoc sp. PCC 7120GP17130610d 0.1 TIGR
- 40. TIGR
- 41. C. pneumoniae Paralogs by Position 1250000 1000000 750000 500000 Subject Orf Position 250000 0 0 250000 500000 750000 1000000 1250000 TIGR Query Orf Position
- 42. C. pneumoniae Paralogs - 1250000 Lineage Specific 1000000 750000 500000 Subject Orf Position 250000 0 0 250000 500000 750000 1000000 1250000 TIGR Query Orf Position