transcription The first step of gene expression – synthesis of RNA molecule Some nomenclature conventions RNAP RNA DNA Similarities and Differences Between DNA and RNA RNA • Similar strand structure • Can define a 5’ and 3’ end • 2’ hydroxyl in RNA: causes stability differences) • Uracil in RNA takes the place of Thymine in in DNA DNA Secondary structure of RNA Stem & loop Difference between DNA and RNA. – DNA – Double Helix, RNA – Single Stranded – DNA --- A-T, RNA ---A-U – DNA ---Deoxyribose Sugar, RNA---Ribose • What is a similarity of DNA and RNA? – G binds with C in both DNA and RNA – Both have sugar and phosphate backbone The pathway of gene expression CAP Conserved seqences Polypyrimidine tract גן הבטא גלובין ההומני Splice sites can be included in the exons mRNA splicing lariat spliceosome מעגל מחזורי של ראקציית השיחבור ספליסוזום Systemic Lupus Erythematosus (SLE) Lupus is a type of immune system disorder known as an autoimmune disease. In autoimmune diseases, the body harms its own healthy cells and tissues. This leads to inflammation and damage of various body tissues. Lupus can affect many parts of the body, including the joints, skin, kidneys, heart, lungs, blood vessels, and brain. Although people with the disease may have many different symptoms, some of the most common ones include extreme fatigue, painful or swollen joints (arthritis), unexplained fever, skin rashes, and kidney problems. Lupus is also known as a rheumatic disease. The rheumatic diseases are a group of disorders that cause aches, pain, and stiffness in the joints, muscles, and bones. 15% of the mutation leading to genetic disorders effect splicing activity Untreated β thalassemia Treatment of thalassemia major antisense Human genome 2.91 billion base pairs 24,500 genes 1.5% exons (127 nucleotides) 24% introns (~3,000 nucleotides) 75% intergenic (no genes) Average size of a gene is 27,894 bases Contains an average of 8.8 exons Titin contains 234 exons. Gene number C. elegans ~19,000 Drosophila melanogaster ~13,600 Arabidopsis thaliana ~25,500 Homo Sapiens ~24,000 Mouse ~24,000 Rice (Oryza sativa) ~50,000 Compared to worm and fly, human has shorter exons and longer introns on the extremes of the distribution 150 nt – drosophila 125 nt – Ciona intes. 119 nt - human 5000 genes 234 introns Multi-introns genes 7.8 introns per gene Sequences of humanmouse homologous exons are 88% conserved Alternative Alternative splicing splicing פוליפרימידין טראק 40% 18% 8% ? 3% isoform 18 איזופורמים 1 Alternative splicing regulate pro or anti apoptotic pathways. pro-apoptotic anti-apoptotic translation The last stage in gene expression – synthesis of protein Amino acid N C N to C terminal 5’ to 3’ CDS Aminoacyl tRNA tRNA Aminoacyl-tRNA • 20( synthetase )diffrent one for each amino – acid NEEDS ENERGY! – 64 combinations for 20 amino acids 3 termination codons סרט טרנסלציה ספירת מלאי Met one codon LEU 6 codons, 4 of which are CUN Disease-Associated Mutations A mutation is a change in the normal base pair sequence Commonly used to define DNA sequence changes that alter protein function Polymorphism DNA sequence changes that do not alter protein function (common definition, not technically correct) Functional protein Functional protein Single nucleotide polymorphisms (SNP) (in the coding sequence) mRNA Normal A U G Protein Sequenc e variant Met A A G U U U GGC GC A U UG C A A Lys Phe Gly Ala Leu Gln mRNA A U G Protein Met A A G U U U GGU GC A U UG C A A Lys Phe Gly Ala Silent DNA sequence polymorphism Leu Gln Missense Mutations Normal mRNA A U G Protein Met A A G U U U GGC GC A U UG C A A Lys Phe Gly Ala Leu Gln mRNA Missense A U G Protein Met A A G U U U AGC GC A U UG C A A Lys Phe Ser Ala Missense: changes to a codon for another amino acid (can be harmful mutation or neutral polymorphism) Leu Gln Nonsense Mutations Normal mRNA A U G Protein Met A A G U U U GGC GC A U UG C A A Lys Phe Gly Ala Leu Gln mRNA A U G Nonsense Protein U A G U U U GGC GC A U UG C A A Met Nonsense: change from an amino acid codon to a stop codon, producing a shortened protein Frameshift Mutations Normal mRNA A U G Protein Frameshift Met A A G U U U GGC GC A U UG C A A Lys Phe Gly Ala Leu Gln mRNA A U G A A G U U G GC G C A U UGC A A Protein Met Lys Leu Ala Frameshift: insertion or deletion of base pairs, producing a stop codon downstream and (usually) shortened protein Pre-mature stop codon mutation Missense mutation Translation II A peptide – a chain of amino acids A protein – a stretch of one or more amino acid chains with biological function +eIF3 eIF3 Translation סרט טרנסלציה Life cycle of mRNA movie