Bioinformatics 1. Sepp Hochreiter. Biology, Sequences, Phylogenetics Part 1. Bioinformatics 1: Biology, Sequences, Phylogenetics
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1 Bioinformatics 1 Biology, Sequences, Phylogenetics Part 1 Sepp Hochreiter
2 3 Credits (plus 3 Credits for exercises) Examination at the end of the class Master Bioinformatics and Computer Science Script is available: separate script - not the power point slides; Slides are available:
3 What is Bioinformatics? Interface of biology and computers Analysis of proteins, genes and genomes using computer algorithms and computer databases Analysis and storage of the billions of DNA base pairs that are sequenced by genomics projects
4 What is Bioinformatics? Computer Science Bioinformatics Biology & Life Sciences Mathematics & Statistics
5 What is Bioinformatics? Bioinformatics is a new subject of genetic data collection, analysis and dissemination to the research community. Hwa A. Lim (1987) Bioinformatics: Research, development, or application of computational tools and approaches for expanding the use of biological, medical, behavioral or health data,including those to acquire, store, organize, archive, analyze, or visualize such data. NIH working definition (2000)
6 Questions Answerd by Bioinformatics From where came the first human? Is Anna Anderson the tsar s daughter Anastasia? Are the neanderthals the ancestors of the humans? What are the evolutionary relationships between species
7 A) from Came B) Not humans from Africa: Africa: competition fromhomo Africa? erectus with homo is human erectus ancestor homo erectus homo sapiens
8 Is Anna Anderson the tsar s daughter Anastasia Romanov? Anastasia (1909) and Anna Anderson (1959) The kids of the tsar
9 Are the neanderthals the human ancestors or a different species?
10 Phylogeny: history of species Phylogenetic knowledge: evolutionary trees turtles birds crocodiles saurians snakes mammals
11 Literature D. W. Mount, Bioinformatics: Sequences and Genome analysis, CSHL Press, 2001 D. Gusfield, Algorithms on strings, trees and sequences: computer science and computational biology, Cambridge Univ. Press, 1999 R. Durbin, S. Eddy, A. Krogh, G. Mitchison, Biological sequence analysis, Cambridge Univ. Press, 1998 M. Waterman, Introduction to Computational Biology, Chapmann & Hall, 1995 Setubal and Meidanis, Introduction to Computational Molecualar Biology, PWS Publishing, 1997 Pevzner, Computational Molecular Biology, MIT Press, 2000 J. Felsenstein: Inferring phylogenies, Sinauer, 2004 W. Ewens, G. Grant, Statistical Methods in Bioinformatics, Springer, 2001 Blast:
12 Contents of Molecular Biology 1.6 Introns, Exons, and Splicing
13 Contents 2 Bioinformatics Rescources 2.1 Data Bases 2.2 Software 2.3 Articles
14 Contents 3 Pairwise Alignment 3.1 Motivation 3.2 Sequence Similarities and Scoring Identity Matrix PAM Matrices BLOSUM Matrices Gap Penalties 3.3 Alignment Algorithms Global Alignment - Needleman-Wunsch Local Alignment - Smith-Waterman Fast Approximations: FASTA, BLAST and BLAT 3.4 Alignment Significance Significance of HSPs Significance of Perfect Matches
15 Contents 4 Multiple Alignment 4.1 Motivation 4.2 Multiple Sequence Similarities and Scoring Consensus and Entropy Score Tree and Star Score Weighted Sum of Pairs Score 4.3 Multiple Alignment Algorithms Exact Methods Progressive Algorithms Other Multiple Alignment Algorithms 4.4 Profiles and Position Specific Scoring Matrices
16 Contents 5 Phylogenetics 5.1 Motivation Tree of Life Molecular Phylogenies Methods 5.2 Maximum Parsimony Methods Tree Length Tree Search Weighted Parsimony and Bootstrapping Inconsistency of Maximum Parsimony 5.3 Distance-based Methods UPGMA Least Squares Minimum Evolution Neighbor Joining Distance Measures 5.4 Maximum Likelihood Methods 5.5 Examples
17 Biological Basics Bioinformatics processes data from molecular biology Molecular biology attempts at discovering the principles of the cell which is the largest unit all lifeforms have in common
18 The Cell
19 The Cell Eukaryotic cells possess a nucleus (plants, vetebrates) Prokaryotic cells do not possess a nucleus (bacteria, archaea)
20 The Cell
21 The Cell
22 The Cell
23 The Cell
24 The Cell
25 Central Dogma How are the nano-machines in the cell constructed? These machines are proteins or protein-rna complexes Where is the information about these machines stored? Everything is stored in the DNA How is the information in the DNA used to build proteins? Central dogma: DNA RNA Proteins
26 Central Dogma 1. transcription (mrna) DNA nucleus 2. transport gene mrna protein trna amino acid chain trna trna 3. translation (ribosom, trna) 4. folding (protein) codons/basetriplets trna ribosom cell membran Amino acid
27 Central Dogma
28 DNA Desoxyribonucleic acid (DNA) codes all information of life double helix as sequence of nucleotides with a deoxyribose ends are called 5' and 3 ; DNA is written from 5' to 3 upstream is towards the 5' end downstream towards the 3' 5 nucleotides (nucleobases, bases): adenine (A), thymine (T), cytosine (C), guanine (G), and uracil (U) first 4 in DNA wheras uracil in RNA instead of thymin two classes: purines (A, G) / pyrimidines (C, U, T)
29 DNA hydrogen bonds between purines and pyrimidines base pairs: A T and C G each helix of the DNA is complementary to the other
30 DNA
31 DNA
32 DNA
33 DNA
34 DNA The DNA is condensed in the nucelus in the chromosomes DNA wraps around histones resulting in chromatin Two chromatins linked at the centromere are a chromosome
35 DNA Single DNA nucleotides differ at each human Small differences are inherited from both parents (except maternal mitochondrial DNA) Variation in the DNA at the same position in at least 1% of the population: single nucleotide polymorphism (SNP -pronounced snip) SNPs occur all 100 to 300 base pairs Current research relate diseases to SNPs (schizophrenia or alcohol dependence).
36 DNA
37 RNA Ribonucleic acid (RNA): sequence of nucleotides Contrast to DNA: ribose rings instead of deoxyribose; uracil instead of thymine transcribed from DNA through RNA polymerases kind of RNA: mrna (messenger), dsrna (double stranded), RNAi (interference), ncrna (non-coding) like trna (codon coding), mirna (micro), sirna (small interfering), rrna (ribosomal)
38 RNA
39 RNA
40 RNA
41 Transcription Transcription is the process of reading out a RNA (mrna) from the DNA
42 Transcription Initiation
43 Transcription Initiation
44 Transcription Initiation
45 Transcription Elongation After 8 nucleotides the sigma-subunit is dissociated from polymerase For elongation there exist promoters
46 Transcription Termination
47 Splicing, Exons and Introns Splicing modifies pre-mrna released after transcription Non-coding sequences: introns (intragenic regions) coding sequences: exons are glued together A snrna complex, the spliceosome, performs the splicing but some RNA sequences can perform autonomous splicing
48 Splicing, Exons and Introns Self-splicing
49 Splicing, Exons and Introns pre-mrna can be spliced in different ways: alternative splicing, therefore a gene can code different proteins Alternative splicing is controlled by signalling molecules
50 Amino Acids
51 Amino Acids
52 Amino Acids Hydrophobic (nonpolar): glycine Gly G methionine alanine Ala A phenylalanine valine Val V tryptophan leucine Leu L proline isoleucine Ile I Hydrophilic (polar) serine Ser S tyrosine threonine Thr T asparagine cysteine Cys C glutamine acidic (-,hydrophilic) aspartic acid Asp D glutamic acid basic (+,hydrophilic) lysine Lys K arginine histidine His H Cysteine and methionine: disulfide bonds Met Phe Trp Pro M F W P Tyr Y Asn N Gln Q Glu E Arg R
53 Amino Acids
54 Genetic Code all proteins consist of these 20 amino acids 3D interactions of the amino acids results in nano-machines genetic code: instructions for producing proteins from DNA protein in coded through a gene which is transcribed into mrna and then translated into an amino acid sequence which automatically configures into a protein genetic code gives the rules for translation rules are simple: 3 nucleotides (codon) = one amino acid AUG and CUG: start codon
55 Genetic Code U U UUU UUC UUA UUG C CUU CUC CUA CUG A AUU AUC AUA AUG G GUU GUC GUA GUG phe leu leu ile met val C A UCU UCC UCA UCG ser UAU UAC UAA UAG pro CAU CAC CAA CAG asn thr AAU AAC AAA AAG ala GAU GAC GAA GAG asp CCU CCC CCA CCG ACU ACC ACA ACG GCU GCC GCA GCG G tyr stop his gln lys glu C = Cytosin, U = Uracil, A = Adenin, G = Guanin Base pairs DNA: A-T and C-G (T = Thymin) UGU UGC UGA UGG CGU CGC CGA CGG AGU AGC AGA AGG GGU GGC GGA GGG cys stop trp arg ser arg gly U C A G U C A G U C A G U C A G
56 Translation After transcription the pre-mrna is spliced, edited, transported out of the nucelus into the cytosol (eukaryotes) The ribosome (protein production machinery) assembles the amina acid sequences out of the mrna Ribosome consists of two subunits 60S and 40S in eukaryotes and 50S and 30S in bacteria
57 Translation Initiation Inactive ribosomes have dissociated subunits Ribosome binds to site at mrna marked by AGGAGGU (Shine-Dalgarno) At this site the initiation factors IF1, IF2, IF3 and 30S ribosomal subunit bind The initiator trna binds to the start codon Then the 50S subunit binds to the complex and translation can start
58 Translation Elongation
59 Translation Elongation
60 Translation Termination Termination by a stop codon (UAA, UAG, UGA) which enters the A-site trnas cannot bind, however release factors bind at or near amio acid chain is released and the 70S ribosome dissoiates 30S subunit remains attached to the mrna and searching for the next Shine-Dalgarno pattern
61 Translation Termination
62 Folding of the Protein Only the correct folded protein functions correctly (cf Creutzfeld-Jacob, Altzheimer, BSE, Parkinson) proteins always fold into their specific 3D structure complicated procedure with lots of interactions folding pathways are not unique and have intermediate states folding is assisted by special chaperones (hide the hydrophobic regions or act as containers) Folding time: miliseconds up to minutes or hours major tasks in bioinformatics is the prediction of the 3D structure to guess the function or to design new proteins
63 Folding of the Protein
64 Folding of the Protein
65 Folding of the Protein
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