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7.2 Transcription & Gene Expression RNA and Protein Synthesis Essential Idea Information stored as a code in DNA is copied onto mRNA. Copyright Pearson Prentice Hall Understanding • Transcription occurs in a 5’ to 3’ direction. • Nucleosomes help to regulate transcription in eukaryotes. • Eukaryotic cells modify mRNA after transcription. • Splicing of mRNA increases the number of different proteins an organism can produce. • Gene expression is regulated by proteins that bind to specific base sequences in DNA. • The environment of a cell and of an organism has an impact on gene expression. Copyright Pearson Prentice Hall IB Assessment Statement State that transcription is carried out in a 5’→ 3’ direction. Copyright Pearson Prentice Hall Direction of Transcription: The 5' end of the nucleotide is added to the 3' of the already existing mRNA chain. http://www.stolaf.edu/people/giannini/flashanimat/molgenetics/transcription.swf Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall The sense strand has the same base sequence as the transcribed mRNA except that the base thymine is replaced by the base uracil. The anti-sense strand acts as the template for the transcription of mRNA. The RNA nucleotides are polymerised along the sugar phosphate backbone by RNA polymerase. Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall IB Assessment Statement Explain the process of transcription in prokaryotes, including the role of the promoter region, RNA polymerase, nucleoside triphosphates and the terminator. http://highered.mcgrawhill.com/sites/9834092339/student_view0/chapter15/mrna_synthesis__transcription_.html Copyright Pearson Prentice Hall Transcription in Prokaryotes • The Promoter region allows the binding of RNA polymerase. The RNA polymerase is then able to: o o o Find the anti-sense strand. Find the start for transcription. Know the direction of transcription. Copyright Pearson Prentice Hall Transcription in Prokaryotes The Promoter region allows the binding of RNA The hydrogen bonds between the bases of the DNA helix are opened up by DNA helicase. The bases of the anti-sense strand ('3 to 5' for DNA) are exposed progressively. RNA nucleotides complementary base pair with the anti-sense nucleotide bases. The free nucleotides (nucleoside triphosphates) are based on RNA. The sugar is the pentose ribose and there are four different nitrogen bases. The nucleotides are adenine, guanine, cytosine and uracil. Copyright Pearson Prentice Hall Transcription in Prokaryotes The RNA polymerase forms covalent bonds between the nucleotides. Free energy is released from the oxidation of the nucleoside triphosphates to form the bond. The bonds are formed by joining the 5' of the free nucleotide to the 3' end of the nucleotide already part of the mRNA chain. The RNA polymerase works along the nucleotides completing the pentosephosphate backbone. Copyright Pearson Prentice Hall Transcription in Prokaryotes The mRNA builds up with the RNA polymerase moving along the anti-sense strand joining the nucleotides. As with the other biochemical processes considered in the syllabus there are additional factor involved in transcription. These are not required for the examination Copyright Pearson Prentice Hall Transcription in Prokaryotes The RNA polymerase reaches the terminator and the RNA polymerase stops. The mRNA is complete Copyright Pearson Prentice Hall Transcription in Prokaryotes Various factors result in the RNA polymerase being released and will return to catalyze another mRNA. The mRNA itself is released from the antis-sense strand. The mRNA strand in prokaryotes can be use straight away unlike the eukaryotic mRNA which requires further modification (see next slide) Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall IB ASSESSMENT STATEMENT State that eukaryotic RNA needs the removal of introns to form mature mRNA Copyright Pearson Prentice Hall RNA Editing mRNA Editing The introns are cut out of RNA molecules. Exon Intron DNA Pre-mRNA The exons are the spliced together to form mRNA. mRNA Cap Tail mRNA Editing re-mRNA has been produced through transcription of the anti-sense strand as described for prokaryotic transcription. (a) The non coding introns are spliced out of the mRNA. The introns are broken down in the nucleus. (b) The remaining mRNA is called mature mRNA and is exported from the nucleus to the cytoplasm for translation into the polypeptide Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall Animation on mRNA editing http://bcs.whfreeman.com/thelifewire/content/chp14/1 401s.swf Copyright Pearson Prentice Hall Translation Messenger RNA is transcribed in the nucleus, and then enters the cytoplasm where it attaches to a ribosome. Nucleus mRNA How are genes regulated? – TWO METHODS 1.Histone proteins in nucleosomes help to regulate transcription in eukaryotes by modifying it structure chemically which enhance or inhibit the expression of genes (transcription) 2.Regulatory Proteins (i.e.hormones)that bind to specific DNA Sequences which enhance or inhibit the expression of genes (transcription) Copyright Pearson Prentice Hall Gene Regulation & Expression Nucleosomes help to regulate transcription in eukaryotes. Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall METHOD 1: Nucleosomes & Gene Expression (the regulation of transcription) • Nucleosomes have proteins called histones. • Chemical modification of these histones are an important factor is determining whether a gene is expressed or NOT. Copyright Pearson Prentice Hall Method 1: Chemical Modification of Histones & Gene expression Types of modifications of histones: 1. Addition of acetyl group to a histone tail 2. Addition of a methyl group to a histone tail Copyright Pearson Prentice Hall Method 1: Chemical Modification of Histones & Gene expression Copyright Pearson Prentice Hall Method 1: Chemical Modification of Histones & Gene expression Chemical Modification of histones can either activate or deactivate genes by increasing or decreasing the accessibility of the gene to transcription factors • Acetyl group: neutralizes the positive charge on histones, making DNA less tightly coiled–> increases transcription • Methyl group: maintains positive charge on histones, making DNA tightly coiled –> decreases transcription Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall Method 1: Chemical Modification of Histones & Gene expression Method 2: Gene expression is regulated by proteins that bind to specific base sequences in DNA. DNA SEQUENCES (in which regulatory proteins can bind) • Enhancers: regulatory sequences on DNA which increase the rate of transcription when proteins bind to them. • Silencer : sequences on DNA which decrease the rate of transcription when proteins bind to them. • Enhancers & Silencers are unique for each gene. Copyright Pearson Prentice Hall Gene Expression by regulatory proteins. https://www.youtube.com/watch?v=KKR28Y _L4CA https://www.youtube.com/watch?v=rs6UkVa OPzo Copyright Pearson Prentice Hall Epigenetics: The environment of a cell and of an organism has an impact on gene expression. http://learn.genetics.utah.edu/content/epigenetic s/intro/ Copyright Pearson Prentice Hall Environmental Affects of Genetic Expression The term epigenetics refers to heritable changes in gene expression (active versus inactive genes) that does NOT involve changes to the underlying DNA sequence; Epigenetic change is a regular and natural occurrence but can also be influenced by several factors: Including age, the environment/lifestyle, and disease state. Copyright Pearson Prentice Hall Environmental Affects of Genetic Expression Epigenetic modifications can manifest as commonly as the manner in which cells terminally differentiate to end up as skin cells, liver cells, brain cells, etc. Or, epigenetic change can have more damaging effects that can result in diseases like cancer. At least TWO systems in which gene expression is modified including: 1. DNA methylation 2. histone modification New and ongoing research is continuously uncovering the role of epigenetics in a variety of human disorders and fatal diseases. Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall END OF SECTION