1. MetaCyc and AraCyc: Plant Metabolic Databases Hartmut Foerster Carnegie Institution

2. Overview  MetaCyc: Explore the Database  AraCyc: The Metabolic Network of Arabidopsis

3. MetaCyc http://www.metacyc.org

4. What is MetaCyc ?  MetaCyc is a multi-organism database that collects any known pathways across all kingdoms  MetaCyc is a curated, literature-based biochemical pathways database  Collaboration between SRI International and Carnegie Institution

5. Goal and Applications Goal Universal repository of metabolic pathways  Up-to-date, literature-curated catalogue of commented enzymes and pathways for use in research, metabolic engineering and education Applications Database of reference used to generate predicted Pathway/Genome DataBases (PGDBs)

6. The Content of MetaCyc  Pathways from primary and secondary (specialized) metabolism Reactions with compound structures  Proteins  Genes Mostly from microorganism and plant kingdoms, and several animal pathways Does not contain sequence information

7. Curation Team  PhD-level curators  Extract data from literature o NLY experimentally verified data ideally protein information  Follow MetaCyc Curator’s guide http://bioinformatics.ai.sri.com/ptools/curatorsguide.pdf

8. MetaCyc Version 10.5 to be released in August, 2006 Note: The statistics for each year pertain to the last MetaCyc version released in that year. Note: The statistics for each year pertain to the last MetaCyc version released in that year

9. MetaCyc – Browse the Database Explore class hierarchy of pathways, compounds, reactions, genes and cell components

10. MetaCyc – Browse the Database (cont’d) Explore class hierarchy of pathways, compounds, reactions, genes and cell components

11. MetaCyc – Explore the Metabolic Universe Query the database (pathways, reactions, compounds, genes)

12. MetaCyc – Query Page Type your search term and click submit Luteolin

13. MetaCyc – Query Result Page

14. MetaCyc – Pathway Detail Page Part I The pathway diagram shows compounds, reactions and metabolic links

15. MetaCyc – Pathway Detail Page Part II Pathway commentary comprises general and specific information about the pathway

16. MetaCyc – More Detail Extend or collapse the detail level of the pathway detail page

17. MetaCyc – More Detail (cont’d) In depth information about reaction, EC number, enzymes, genes, regulatory aspects, and metabolic links to related pathways

18. MetaCyc – More Detail (cont’d) More detail reveals structural information about compounds

19. MetaCyc – Reaction Detail Page

20. MetaCyc – Enzymes and Genes Contains enzyme commentary, references, and physico-chemical properties of the enzyme

21. MetaCyc – Enzymes and Genes (cont’d)

22. MetaCyc – Evidence Codes Intuitive icons Pathway Level Evidence codes provide assessment of data quality, i.e. the affirmation for the existence of an pathway Evidence codes provide assessment of data quality, i.e. the affirmation for the catalytic activity of an enzyme Enzyme Level

23. MetaCyc – Evidence Codes (cont’d)

24. Variants, Related Pathways and Links  Pathway variants are created as separate pathways  IAA biosynthesis I (tryptophane-dependent) IAA biosynthesis II (tryptophane-independent)  Links are added between interconnected pathways  Related pathways are grouped into superpathways  e.g.superpathway of choline biosynthesis

25. Creation of a Superpathway L-serine ethanolamine phosphoryl-ethanolamine N-methylethanolamine phosphate N-dimethylethanolamine phosphate phosphoryl-choline choline biosynthesis III Choline biosynthesis II ethanolamine phosphoryl-choline CDP-choline-choline a phosphatidylcholine choline biosynthesis IIIcholine biosynthesis IIcholine biosynthesis I N-monomethylethanolamine N-dimethylethanolamine choline Superpathway choline biosynthesis

26. Applications: Pathways Prediction Goal Universal repository of metabolic pathways  Up-to-date, literature-curated catalogue of commented enzymes and pathways for use in research, metabolic engineering and education Applications Database of reference used to generate predicted Pathway/Genome DataBases (PGDBs)

27. Pathway Tools Software Suite  Software for generating, curating, querying, displaying PGDBs  Developed by Peter Karp and team PathoLogic – Infers pathways from genome or transcripts sequencing Pathway/Genome Editors – Curation interface Pathway/Genome Navigator – Query, visualization, analysis and Web publishing OMICS Viewer

28. The PGDB Family Annotated Genome Arabidopsis thaliana PathoLogic Software Reference Pathway Database (MetaCyc) Reactions Pathways compounds Gene products genes Pathway/Genome Database (AraCyc)

29. AraCyc: The Arabidopsis thaliana specific metabolic database http://www.arabidopsis.org/tools/aracyc

30. The Computational Build of AraCyc  In 2004, the Arabidopsis genome contained 7900 genes annotated to the GO term ‘catalytic activity’  4900 enzymes in small molecule metabolism (19% of the total genome)  PathoLogic inferred 219 pathways and mapped 940 (19% enzyme-coding) genes to the pathways

31. Cleaning of a Newborn Database  PathoLogic errs on the side of over-prediction  First round of curation to remove false-positives  Add missing pathways  Improve the quality of information  Introduce new pathways  Increase number of pathway and protein comments  Refine computational assignment of protein

32. Pathway Validation Criteria  A pathway that is described in the Arabidopsis literature  A pathway whose critical metabolites are described in the Arabidopsis literature  A pathway that contains unique reactions and having genes assigned to those unique reactions

33. Validation Procedure  Delete non-plant pathways: Pathway variants of bacteria-origin Pathways not operating in plants at all (e.g. glycogen biosynthesis)  Add new plant-specific pathways: Pathway variants of plant-origin Plant-specific metabolites (e.g. plant hormones) Plant-specific metabolism (e.g. xanthophyll cycle)

34. AraCyc - Curation Progress AraCyc (2.1) Apr 2005 AraCyc (2.5) Oct 2005 AraCyc (2.6) Feb 2006 Total pathways221197228 New-3735 Updated-04 Deleted-616 Pathways manually reviewed with literature evidence 71 (32%)170 (86%)201 (88%) Additional 66 new pathways were added to MetaCyc

35. How to Link to AraCyc From the TAIR home page click on the link to AraCyc pathways

36. AraCyc – The Home Page Browse pathways, enzymes, genes, compounds Display of the Arabidopsis metabolic network Paint data from high-throughput experiments on the metabolic map User submission form

37. AraCyc – All the Help you can get

38. AraCyc’s Content

39. Computationally Predicted Enzymes

40. Experimentally Verified Enzymes

41. AraCyc: Pathway Detail Page (cont’d)

42. AraCyc: OmicsViewer

43. OMICS Viewer  Part of the Pathway Tools Software Suite  Displays bird-eye view of the Metabolic Overview diagram  Allows to paint data values from the user's high- throughput onto the Metabolic Overview diagram  Microarray Expression Data  Proteomics Data  Metabolomics Data

44. OmicsViewer Submission Page Step 1 Step 2 Load sample file and provide information about your data Sample data file (text tab-delimited) 0 1 2 3 4

45. OmicsViewer Submission Page (cont’d) Step 3 Step 6 Step 4 Step 5 Choose relative or absolute values Check the box if you have log values or negative fold change numbers Choose to display a single or multiple step experiment Select the type of data you want to display (refers to your loading file)

46. OmicsViewer Submission Page (cont’d) Step 7 For single or multiple time points add the corresponding column number(s)

47. Step 8 Step 9 OmicsViewer Submission Page (cont’d) Choose your cutoff to visualize your expression values

48. The Omics Viewer Result Page reactions (lines) are color-coded according to the gene expression level compounds (icons) are color-coded according to the concentration of compounds

49. The Omics Viewer Result Page (cont’d) The statistics for the expression map (single time points only) is provided at the bottom of the page

51. Saving Results

52. Curation Flow

53. Acknowledgements  TAIR - Sue Rhee (PI) - Peifen Zhang (curator) - Christophe Tissier (curator) - Hartmut Foerster (curator) - Mary Montoya (software developer) - Joe Filla (sysadmin)  SRI - Peter Karp (PI) - Ron Caspi (curator) - Carol Fulcher (curator) - Suzanne Paley (software developer) - Pallavi Kaipa (programmer) - Markus Krummenacker (programmer) - Mario Latendresse (programmer) NIH,NSF,Pioneer Hi-Bred  Previous contributers - Lukas Müller - Aleksey Kleytman (curator assistant – TAIR) - Thomas Yan (programmer – TAIR) - John Pick (software developer - SRI)