Module 7: The Central Dogma. CSE590: Molecular programming and neural computation. All slides by Eric Klavins.
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1 Module 7: The Central Dogma CSE590: Molecular programming and neural computation. All slides by Eric Klavins. 1
2 The Central Dogma RNA Polymerase The Ribosome m Note: We will look mainly at prokaryo=c (e.g. e. coli) processes. Some of this is the same in eukaryotes, but there are important differences.
3 Important Molecules DNA = Deoxyribonucleic acid A sequence of A, T, C and G (deoxyribonucleo=des) RNA = Ribonucleic Acid (mrna, trna, ) A sequence of A, U, C and G (ribonucleo=des) RNA Polymerase (RNAP) Transcribes (copies) DNA segments into RNA Amino Acids and Transfer RNA (trna) Help build proteins The Ribosome Translates messenger RNA (mrna) into protein Protein A sequence of amino acids
4 5 end 3 end Nucleo=des
5 DNA Nucleo=des can make single stranded polymers called ssdna (single stranded DNA). Complementary ssdnas can hybridize to form dsdna (double stranded DNA).
6 Drawing DNA You have to say which way the DNA goes, usually from 5 to 3. A single stranded DNA: 5 -ATCCTGTAATGC-3 A double stranded DNA: 5 -ATCCTGTAATGC-3 3 -TAGGACATTACG-5 Some=mes, we write dsdna by wri=ng only one strand, since the other is implied, as in genebank data.
7 2nm The double helix 10 base pairs per turn: 3.4 nm Minor groove Major groove The informa=on of DNA is encoded in the sequence of bases. The sequence can also effect the detailed structure of the major and minor grooves. Repressor and promotor proteins can bind with high specificity to the outside of the DNA helix (more on this next week).
8 DNA is long! E. Coli genome = 4.6 Mbp (one molecule!) human genome = 2m long! DNA is stored under pressure in virus capsids. This 160,000 bp piece of DNA exploded from within a T4 bacteriophage.
9 Important Molecules DNA = Deoxyribonucleic acid A sequence of A, T, C and G (deoxyribonucleo=des) RNA = Ribonucleic Acid (mrna, trna, ) A sequence of A, U, C and G (ribonucleo=des) RNA Polymerase (RNAP) Transcribes (copies) DNA segments into RNA Amino Acids and Transfer RNA (trna) Help build proteins The Ribosome Translates messenger RNA (mrna) into protein Protein A sequence of amino acids
10 DNA is long. RNA is short. DNA is stable. RNAs appear and disappear. DNA is double stranded. RNA is single stranded and somebmes folds up on itself in funny ways. DNA stores informa=on. RNA does many things!
11 Important Molecules DNA = Deoxyribonucleic acid A sequence of A, T, C and G (deoxyribonucleo=des) RNA = Ribonucleic Acid (mrna, trna, ) A sequence of A, U, C and G (ribonucleo=des) RNA Polymerase (RNAP) Transcribes (copies) DNA segments into RNA Amino Acids and Transfer RNA (trna) Help build proteins The Ribosome Translates messenger RNA (mrna) into protein Protein A sequence of amino acids
12 RNA is transcribed from DNA templates triphoshpate The region that expresses an RNA is called a gene. Most mrnas are 3000 bp or, usually, shorter.
13 All types of RNA are generated this way Transcrip1on mrna Sometimes several mrnas are copied in a row to produce a string of protein coding mrnas. trna (e. coli has ~60 trna genes) sirna Eukaryotes microrna Ribosomal RNA (e. coli has 7 copies of its rrna genes so that it can produce a lot of ribosomes) RNA is totally the coolest molecule.
14 RNA Polymerase (RNAP) The workhorse of transcrip=on is RNAP. RNAP catalyzes the forma=on of the phosphodiester bond linking the nucleo=des together in RNA. RNAP moves at about 20 nucleo=des per second! Some genes are transcribed faster than others. RNAP is a holoenzyme, consis=ng of 4 proteins subunits and a detachable cofactor enzyme (σ- cofactor). If the wrong nucleo=de is added, RNAP backs up and fixes its mistakes!
15 Start signals are coded in DNA Promotor - 35 TTGACAT - 10 TATAAT 1 downstream upstream 1. RNAP slides along DNA without transcribing. 2. σ-factor binds to promotor 3. RNAP binds with σ- factor un=l about 10 bases are transcribed. 4. When you write DNA programs, these are the things you specify!
16 Stop signals are encoded in DNA Terminators are AAAAAA sequences preceded by a short palindrome. The palindrome forms a hairpin in the growing RNA. The shape of the hairpined RNA pops RNAP off the gene and transcrip=on stops!
17 Important Molecules DNA = Deoxyribonucleic acid A sequence of A, T, C and G (deoxyribonucleo=des) RNA = Ribonucleic Acid (mrna, trna, ) A sequence of A, U, C and G (ribonucleo=des) RNA Polymerase (RNAP) Transcribes (copies) DNA segments into RNA Amino Acids and Transfer RNA (trna) Help build proteins The Ribosome Translates messenger RNA (mrna) into protein Protein A sequence of amino acids
18 Amino Acids Amino Carboxyl Group One of 20 residues c terminus n terminus Amino acids condense to form chains called polypep1des.
19 The 20 Amino Acids Each one has a different shape, charge, and hydrophobcity. By linking up some into a long chain, you can make a liele machine. We know how to make some kinds of machines this way, but for the most part we don t know much about how to do this in a principled manner.
20 Transfer RNA (trna) Each amino acid can have several trnas, one for each codon varia=on. Various trna synthetases and other enzymes provide a post- transla=onal modifica=on that adds the amino acid.
21 Proteins grow at the carboxyl group (C- terminus) C- Terminus
22 RNA is translated to protein Each 3 nucleo=de sequence, called a codon, in mrna (from 5 to 3 ) codes for an amino acid. Transla=on is the process of building the corresponding protein from this code. 5 - CUC AGC GUU ACC- 3 Leu Ser Val Thr
23 The start and stop codons Transla=on starts with the codon AUG. So all proteins start with methionine. Transla=on stops with UAA, UAG or UGA. These do not code for amino acids.
24 Important Molecules DNA = Deoxyribonucleic acid A sequence of A, T, C and G (deoxyribonucleo=des) RNA = Ribonucleic Acid (mrna, trna, ) A sequence of A, U, C and G (ribonucleo=des) RNA Polymerase (RNAP) Transcribes (copies) DNA segments into RNA Amino Acids and Transfer RNA (trna) Help build proteins The Ribosome Translates messenger RNA (mrna) into protein Protein A sequence of amino acids
25 The Ribosome Binding Site Ribosomes bind to specific sequences on mrnas to ini=ate transla=on. +1 Coding O D - 35 O C - 10 O P RBS Seq 5 UTR Shine- Dalgarno region Spacer Start codon nt Shine- Dalgarno region docks 16S rrna Start codon, first 10+ codons affect binding affinity RBS spacer length modulates transla=on ini=a=on rate
26 Ribosomes translate mrna to protein The two subunits of the ribosome are separate un=l transla=on starts. In bacteria, a ribosome processes 20 amino acids per second. The ribosome makes one mistake every amino acids
27 The ribosome (2/3 RNA, 1/3 Protein)
28 The RNA World There are those who believe that RNA was the first autocataly=c molecular system. RNA can cleave and ligate itself. RNAs have been designed that can transcribe RNA. The ribosome is made almost en=rely out of RNA. Protein and DNA may have come along later. Having separate molecules for informa=on and structure may have been evolu=onarily advantageous.
29 Important Molecules DNA = Deoxyribonucleic acid A sequence of A, T, C and G (deoxyribonucleo=des) RNA = Ribonucleic Acid (mrna, trna, ) A sequence of A, U, C and G (ribonucleo=des) RNA Polymerase (RNAP) Transcribes (copies) DNA segments into RNA Amino Acids and Transfer RNA (trna) Help build proteins The Ribosome Translates messenger RNA (mrna) into protein Protein A sequence of amino acids
30 Proteins Ball and s=ck Structure cartoon Space filling Primary structure: The sequence of amino acids Secondary structure: The local shape (helix, coil or sheet) Ter=ary structure: The global 3D shape Quaternary structure: How proteins form groups
31 Proteins form groups Hemoglobin consists of four protein subunits and four non- protein iron containing heme units. It is self- assembled inside the cell once the components are present.
32 Proteins can be Enzymes An enzyme is a proteins that accelerates a chemical reac=on, usually very specifically.
33 Proteins can pump small molecules (2003 Nobel Prize to Peter Agre) A 1 nanosecond simula=on of the 60,000 atom aquaporin- 1 water channel with full electrosta=cs and constant pressure in a single week (Schulten Group, UIUC). Aquaporin transmembrane four- protein complex. Phospholipid membrane
34 Proteins are involved with signaling
35 RNA and Protein Are Degraded RNA is degraded by Ribonucleases (Rnase) Protein is degraded by Proteases Some RNAs and Proteins are more stable than others. Synthe=c Biologists can tune degrada=on rates.
36 Important Molecules DNA = Deoxyribonucleic acid A sequence of A, T, C and G (deoxyribonucleo=des) RNA = Ribonucleic Acid (mrna, trna, ) A sequence of A, U, C and G (ribonucleo=des) RNA Polymerase (RNAP) Transcribes (copies) DNA segments into RNA Amino Acids and Transfer RNA (trna) Help build proteins The Ribosome Translates messenger RNA (mrna) into protein Protein A sequence of amino acids
37 In prokaryotes, everything happens at once!
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