1. HIDDEN MARKOV MODELS IN MULTIPLE ALIGNMENT

2. 2 HMM Architecture Markov Chains What is a Hidden Markov Model(HMM)? Components of HMM Problems of HMMs

3. 3 Markov Chains SunnyRainCloudy States : Three states - sunny, cloudy, rainy. Time (days)

4. 4 Markov Chains Initial Distribution : Defining the probability of the system being in each of the states at time 0. State transition matrix : The probability of the weather given the previous day's weather. Sunny Rain Cloudy States : Three states - sunny, cloudy, rainy.

5. 5 Hidden Markov Models Hidden states : the (TRUE) states of a system that may be described by a Markov process (e.g., High of low pressure systems). Observable states : the states of the process that are `visible' (e.g., weather). HHHLLL

6. 6 Components Of HMM Output matrix : containing the probability of observing a particular observable state given that the hidden model is in a particular hidden state. Initial Distribution : contains the probability of the (hidden) model being in a particular hidden state at time t = 1. State transition matrix : holding the probability of a hidden state given the previous hidden state.

7. 7 Protein Profile HMMs Motivation –Given a single amino acid target sequence of unknown structure, we want to infer the structure of the resulting protein. Use Profile Similarity What is a Profile? –Proteins families of related sequences and structures –Same function –Clear evolutionary relationship –Patterns of conservation, some positions are more conserved than the others

8. 8 Aligned Sequences Build a Profile HMM (Training) Database search Multiple alignments (Viterbi) Query against Profile HMM database (Forward) An Overview

9. 9 Matching states Insertion states Deletion states No of matching states = average sequence length in the family PFAM Database - of Protein families (http://pfam.wustl.edu)http://pfam.wustl.edu HMM topology

10. A HMM model for a DNA motif alignments, The transitions are shown with arrows whose thickness indicate their probability. In each state, the histogram shows the probabilities of the four bases. ACA - - - ATG TCA ACT ATC ACA C - - AGC AGA - - - ATC ACC G - - ATC Building – from an existing alignment Transition probabilities Output Probabilities insertion

11. 11 Matching states Insertion states Deletion states No of matching states = average sequence length in the family PFAM Database - of Protein families (http://pfam.wustl.edu)http://pfam.wustl.edu Building – Final Topology

12. 12 Given HMM, M, for a sequence family, find all members of the family in data base. Database Searching

13. Consensus sequence: P (ACACATC) = 0.8x1 x 0.8x1 x 0.8x0.6 x 0.4x0.6 x 1x1 x 0.8x1 x 0.8 = 4.7 x 10 -2 Suppose I have a query protein sequence, and I am interested in which family it belongs to? There can be many paths leading to the generation of this sequence. Need to find all these paths and sum the probabilities. ACAC - - ATC Query a new sequence

14. 14 Multiple Alignments Try every possible path through the model that would produce the target sequences –Keep the best one and its probability. –Output : Sequence of match, insert and delete states Viterbi alg. Dynamic Programming

15. 15 PHMM Example An alignment of 30 short amino acid sequences chopped out of a alignment of the SH3 domain. The shaded area are the most conserved and were represented by the main states in the HMM. The unshaded area was represented by an insert state.SH3 domain

16. 16 Advantages Characterize an entire family of sequences. Position-dependent character distributions and position-dependent insertion and deletion gap penalties. Built on a formal probabilistic basis Can make libraries of hundreds of profile HMMs and apply them on a large scale (whole genome)

17. 17 Limitations Markov Chains Probabilities of states are supposed to be independent P(y) must be independent of P(x), and vice versa This usually isn’t true P(x) … P(y)