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Inferring Function from Homology

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Bioinformatics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1526))

Abstract

Recent technological advances in sequencing and high-throughput DNA cloning have resulted in the generation of vast quantities of biological sequence data. Ideally the functions of individual genes and proteins predicted by these methods should be assessed experimentally within the context of a defined hypothesis. However, if no hypothesis is known a priori, or the number of sequences to be assessed is large, bioinformatics techniques may be useful in predicting function.

This chapter proposes a pipeline of freely available Web-based tools to analyze protein-coding DNA and peptide sequences of unknown function. Accumulated information obtained during each step of the pipeline is used to build a testable hypothesis of function.

The following methods are described in detail:

  1. 1.

    Annotation of gene function through Protein domain detection (SMART and Pfam).

  2. 2.

    Sequence similarity methods for homolog detection (BLAST and DELTA-BLAST).

  3. 3.

    Comparing sequences to whole genome data.

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References

  1. Doolittle RF (1981) Similar amino acid sequences: chance or common ancestry? Science 214(4517):149–159

    Article  CAS  PubMed  Google Scholar 

  2. Pearson WR, Sierk ML (2005) The limits of protein sequence comparison? Curr Opin Struct Biol 15(3):254–260

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Fitch WM (2000) Homology a personal view on some of the problems. Trends Genet 16(5):227–231

    Article  CAS  PubMed  Google Scholar 

  4. Henikoff S, Greene EA, Pietrokovski S, Bork P, Attwood TK, Hood L (1997) Gene families: the taxonomy of protein paralogs and chimeras. Science 278(5338):609–614

    Article  CAS  PubMed  Google Scholar 

  5. Sonnhammer EL, Koonin EV (2002) Orthology, paralogy and proposed classification for paralog subtypes. Trends Genet 18(12):619–620

    Article  CAS  PubMed  Google Scholar 

  6. Weber MJ (2005) New human and mouse microRNA genes found by homology search. FEBS J 272(1):59–73

    Article  CAS  PubMed  Google Scholar 

  7. Tatusov RL, Galperin MY, Natale DA, Koonin EV (2000) The COG database: a tool for genome-scale analysis of protein functions and evolution. Nucleic Acids Res 28(1):33–36

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Hurles M (2004) Gene duplication: the genomic trade in spare parts. PLoS Biol 2(7):E206

    Article  PubMed  PubMed Central  Google Scholar 

  9. Bateman A (1997) The structure of a domain common to archaebacteria and the homocystinuria disease protein. Trends Biochem Sci 22(1):12–13

    Article  CAS  PubMed  Google Scholar 

  10. Ponting CP, Russell RR (2002) The natural history of protein domains. Annu Rev Biophys Biomol Struct 31:45–71

    Article  CAS  PubMed  Google Scholar 

  11. Ponting CP (2001) Issues in predicting protein function from sequence. Brief Bioinform 2(1):19–29

    Article  CAS  PubMed  Google Scholar 

  12. Ponting CP, Dickens NJ (2001) Genome cartography through domain annotation. Genome Biol 2(7), Comment 2006

    Google Scholar 

  13. Flicek P, Ahmed I, Amode MR, Barrell D, Beal K, Brent S, Carvalho-Silva D, Clapham P, Coates G, Fairley S et al (2013) Ensembl 2013. Nucleic Acids Res 41(Database issue):D48–D55

    Article  CAS  PubMed  Google Scholar 

  14. Hubbard T, Barker D, Birney E, Cameron G, Chen Y, Clark L, Cox T, Cuff J, Curwen V, Down T et al (2002) The Ensembl genome database project. Nucleic Acids Res 30(1):38–41

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Meyer LR, Zweig AS, Hinrichs AS, Karolchik D, Kuhn RM, Wong M, Sloan CA, Rosenbloom KR, Roe G, Rhead B et al (2013) The UCSC Genome Browser database: extensions and updates 2013. Nucleic Acids Res 41(Database issue):D64–D69

    Article  CAS  PubMed  Google Scholar 

  16. Marchler-Bauer A, Panchenko AR, Shoemaker BA, Thiessen PA, Geer LY, Bryant SH (2002) CDD: a database of conserved domain alignments with links to domain three-dimensional structure. Nucleic Acids Res 30(1):281–283

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Marchler-Bauer A, Zheng C, Chitsaz F, Derbyshire MK, Geer LY, Geer RC, Gonzales NR, Gwadz M, Hurwitz DI, Lanczycki CJ, Lu F, Lu S, Marchler GH, Song JS, Thanki N, Yamashita RA, Zhang D, Bryant SH (2013) CDD: conserved domains and protein three-dimensional structure. Nucleic Acids Res 41(Database issue):D348–D352

    Article  CAS  PubMed  Google Scholar 

  18. Apweiler R, Attwood TK, Bairoch A, Bateman A, Birney E, Biswas M, Bucher P, Cerutti L, Corpet F, Croning MD et al (2001) The InterPro database, an integrated documentation resource for protein families, domains and functional sites. Nucleic Acids Res 29(1):37–40

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Jones P, Binns D, Chang HY, Fraser M, Li W, McAnulla C, McWilliam H, Maslen J, Mitchell A, Nuka G, Pesseat S, Quinn AF, Sangrador-Vegas A, Scheremetjew M, Yong SY, Lopez R, Hunter S (2014) InterProScan 5: genome-scale protein function classification. Bioinformatics 30(9):1236–1240

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Bateman A, Birney E, Durbin R, Eddy SR, Finn RD, Sonnhammer EL (1999) Pfam 3.1: 1313 multiple alignments and profile HMMs match the majority of proteins. Nucleic Acids Res 27(1):260–262

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Finn RD, Bateman A, Clements J, Coggill P, Eberhardt RY, Eddy SR, Heger A, Hetherington K, Holm L, Mistry J, Sonnhammer EL, Tate J, Punta M (2014) Pfam: the protein families database. Nucleic Acids Res 42(Database issue):D222–D230

    Article  CAS  PubMed  Google Scholar 

  22. Letunic I, Doerks T, Bork P (2012) SMART 7: recent updates to the protein domain annotation resource. Nucleic Acids Res 40(Database issue):D302–D305

    Article  CAS  PubMed  Google Scholar 

  23. Schultz J, Milpetz F, Bork P, Ponting CP (1998) SMART, a simple modular architecture research tool: identification of signaling domains. Proc Natl Acad Sci U S A 95(11):5857–5864

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A 85(8):2444–2448

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215(3):403–410

    Article  CAS  PubMed  Google Scholar 

  26. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25(17):3389–3402

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Eddy SR (1998) Profile hidden Markov models. Bioinformatics 14(9):755–763

    Article  CAS  PubMed  Google Scholar 

  28. Gibbs RA, Weinstock GM, Metzker ML, Muzny DM, Sodergren EJ, Scherer S, Scott G, Steffen D, Worley KC, Burch PE et al (2004) Genome sequence of the Brown Norway rat yields insights into mammalian evolution. Nature 428(6982):493–521

    Article  CAS  PubMed  Google Scholar 

  29. Lander ES, Linton LM, Birren B, Nusbaum C, Zody MC, Baldwin J, Devon K, Dewar K, Doyle M, FitzHugh W et al (2001) Initial sequencing and analysis of the human genome. Nature 409(6822):860–921

    Article  CAS  PubMed  Google Scholar 

  30. Ellsworth RE, Jamison DC, Touchman JW, Chissoe SL, Braden Maduro VV, Bouffard GG, Dietrich NL, Beckstrom-Sternberg SM, Iyer LM, Weintraub LA et al (2000) Comparative genomic sequence analysis of the human and mouse cystic fibrosis transmembrane conductance regulator genes. Proc Natl Acad Sci U S A 97(3):1172–1177

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Emes RD, Goodstadt L, Winter EE, Ponting CP (2003) Comparison of the genomes of human and mouse lays the foundation of genome zoology. Hum Mol Genet 12(7):701–709

    Article  CAS  PubMed  Google Scholar 

  32. Schultz J, Copley RR, Doerks T, Ponting CP, Bork P (2000) SMART: a web-based tool for the study of genetically mobile domains. Nucleic Acids Res 28(1):231–234

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Sonnhammer EL, Eddy SR, Birney E, Bateman A, Durbin R (1998) Pfam: multiple sequence alignments and HMM-profiles of protein domains. Nucleic Acids Res 26(1):320–322

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Finn RD, Mistry J, Schuster-Bockler B, Griffiths-Jones S, Hollich V, Lassmann T, Moxon S, Marshall M, Khanna A, Durbin R, Eddy SR, Sonnhammer EL, Bateman A (2006) Pfam: clans, web tools and services. Nucleic Acids Res 34(Database issue):D247–D251

    Article  CAS  PubMed  Google Scholar 

  35. Henikoff S, Henikoff JG (1993) Performance evaluation of amino acid substitution matrices. Proteins 17(1):49–61

    Article  CAS  PubMed  Google Scholar 

  36. Henikoff S, Henikoff JG (1992) Amino acid substitution matrices from protein blocks. Proc Natl Acad Sci U S A 89(22):10915–10919

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Smith TF, Waterman MS (1981) Identification of common molecular subsequences. J Mol Biol 147(1):195–197

    Article  CAS  PubMed  Google Scholar 

  38. Wheeler DL, Barrett T, Benson DA, Bryant SH, Canese K, Chetvernin V, Church DM, Dicuccio M, Edgar R, Federhen S et al (2008) Database resources of the National Center for Biotechnology Information. Nucleic Acids Res 36(Database issue):D13–D21

    CAS  PubMed  Google Scholar 

  39. Pearson WR (2014) BLAST and FASTA similarity searching for multiple sequence alignment. Methods Mol Biol 1079:75–101

    Article  PubMed  Google Scholar 

  40. Altschul SF, Gertz EM, Agarwala R, Schaffer AA, Yu YK (2009) PSI-BLAST pseudocounts and the minimum description length principle. Nucleic Acids Res 37(3):815–824

    Article  CAS  PubMed  Google Scholar 

  41. Altschul SF, Koonin EV (1998) Iterated profile searches with PSI-BLAST—a tool for discovery in protein databases. Trends Biochem Sci 23(11):444–447

    Article  CAS  PubMed  Google Scholar 

  42. Boratyn GM, Schaffer AA, Agarwala R, Altschul SF, Lipman DJ, Madden TL (2012) Domain enhanced lookup time accelerated BLAST. Biol Direct 7:12

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Jones DT, Swindells MB (2002) Getting the most from PSI-BLAST. Trends Biochem Sci 27(3):161–164

    Article  CAS  PubMed  Google Scholar 

  44. Korf I (2003) Serial BLAST searching. Bioinformatics 19(12):1492–1496

    Article  CAS  PubMed  Google Scholar 

  45. Altschul SF, Bundschuh R, Olsen R, Hwa T (2001) The estimation of statistical parameters for local alignment score distributions. Nucleic Acids Res 29(2):351–361

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Wootton JC, Federhen S (1996) Analysis of compositionally biased regions in sequence databases. Methods Enzymol 266:554–571

    Article  CAS  PubMed  Google Scholar 

  47. Altschul SF, Gish W (1996) Local alignment statistics. Methods Enzymol 266:460–480

    Article  CAS  PubMed  Google Scholar 

  48. Henikoff S (1996) Scores for sequence searches and alignments. Curr Opin Struct Biol 6(3):353–360

    Article  CAS  PubMed  Google Scholar 

  49. Schaffer AA, Aravind L, Madden TL, Shavirin S, Spouge JL, Wolf YI, Koonin EV, Altschul SF (2001) Improving the accuracy of PSI-BLAST protein database searches with composition-based statistics and other refinements. Nucleic Acids Res 29(14):2994–3005

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Sierk ML, Pearson WR (2004) Sensitivity and selectivity in protein structure comparison. Protein Sci 13(3):773–785

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Wass MN, Barton G, Sternberg MJ (2012) CombFunc: predicting protein function using heterogeneous data sources. Nucleic Acids Res 40(Web Server issue):W466–W470

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Minneci F, Piovesan D, Cozzetto D, Jones DT (2013) FFPred 2.0: improved homology-independent prediction of gene ontology terms for eukaryotic protein sequences. PLoS One 8(5):e63754

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Ashburner M, Ball CA, Blake JA, Botstein D, Butler H, Cherry JM, Davis AP, Dolinski K, Dwight SS, Eppig JT et al (2000) Gene ontology: tool for the unification of biology. The Gene Ontology Consortium. Nat Genet 25(1):25–29

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Henikoff S, Pietrokovski S, Henikoff JG (1998) Superior performance in protein homology detection with the Blocks Database servers. Nucleic Acids Res 26(1):309–312

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Henikoff JG, Pietrokovski S, McCallum CM, Henikoff S (2000) Blocks-based methods for detecting protein homology. Electrophoresis 21(9):1700–1706

    Article  CAS  PubMed  Google Scholar 

  56. Schaffer AA, Wolf YI, Ponting CP, Koonin EV, Aravind L, Altschul SF (1999) IMPALA: matching a protein sequence against a collection of PSI-BLAST-constructed position-specific score matrices. Bioinformatics 15(12):1000–1011

    Article  CAS  PubMed  Google Scholar 

  57. Pietrokovski S (1996) Searching databases of conserved sequence regions by aligning protein multiple-alignments. Nucleic Acids Res 24(19):3836–3845

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Sadreyev R, Grishin N (2003) COMPASS: a tool for comparison of multiple protein alignments with assessment of statistical significance. J Mol Biol 326(1):317–336

    Article  CAS  PubMed  Google Scholar 

  59. Sadreyev RI, Grishin NV (2004) Quality of alignment comparison by COMPASS improves with inclusion of diverse confident homologs. Bioinformatics 20(6):818–828

    Article  CAS  PubMed  Google Scholar 

  60. Sadreyev RI, Tang M, Kim BH, Grishin NV (2007) COMPASS server for remote homology inference. Nucleic Acids Res 35(Web Server issue):W653–W658

    Article  PubMed  PubMed Central  Google Scholar 

  61. Sadreyev RI, Tang M, Kim BH, Grishin NV (2009) COMPASS server for homology detection: improved statistical accuracy, speed and functionality. Nucleic Acids Res 37(Web Server issue):W90–W94

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. Soding J, Biegert A, Lupas AN (2005) The HHpred interactive server for protein homology detection and structure prediction. Nucleic Acids Res 33(Web Server issue):W244–W248

    Article  PubMed  PubMed Central  Google Scholar 

  63. Hildebrand A, Remmert M, Biegert A, Soding J (2009) Fast and accurate automatic structure prediction with HHpred. Proteins 77(Suppl 9):128–132

    Article  CAS  PubMed  Google Scholar 

  64. Eddy SR (2011) Accelerated profile HMM searches. PLoS Comput Biol 7(10):e1002195

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. Kent WJ (2002) BLAT—the BLAST-like alignment tool. Genome Res 12(4):656–664

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  66. Marchler-Bauer A, Anderson JB, Derbyshire MK, DeWeese-Scott C, Gonzales NR, Gwadz M, Hao L, He S, Hurwitz DI, Jackson JD et al (2007) CDD: a conserved domain database for interactive domain family analysis. Nucleic Acids Res 35(Database issue):D237–D240

    Article  CAS  PubMed  Google Scholar 

  67. Wheeler DL, Church DM, Federhen S, Lash AE, Madden TL, Pontius JU, Schuler GD, Schriml LM, Sequeira E, Tatusova TA, Wagner L (2003) Database resources of the National Center for Biotechnology. Nucleic Acids Res 31(1):28–33

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Correspondence to Richard D. Emes .

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Giles, T.C., Emes, R.D. (2017). Inferring Function from Homology. In: Keith, J. (eds) Bioinformatics. Methods in Molecular Biology, vol 1526. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6613-4_2

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  • DOI: https://doi.org/10.1007/978-1-4939-6613-4_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6611-0

  • Online ISBN: 978-1-4939-6613-4

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