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Gene to Protein Chapter 17 Chapter 17 Central Dogma of Molecular Biology Chapter 16 Chapter 17 Chapter 17 Overview of Central Dogma Transcription: The process of building an mRNA strand from DNA. Translation: The process of building a polypeptide from an mRNA transcript. Ultimate goal: Use a sequence of DNA to get a protein. Chapter 17 DNA mRNA Polypeptide Transcription: DNA mRNA occurs in nucleus Translation: mRNA Polypeptide occurs in cytoplasm or RER Chapter 17 Transcription DNA RNA Much like replication with one major difference: C goes with G A goes with U (Not T!) GATAGC TACAG TGGGAGT ATGCG T A TC TCC CUAUCGAUGUCACCC UCAUACGCAUAGAGG Chapter 17 The Genetic Code Codon: A three-nucleotide sequence encoding one amino acid Eg – ACA (adenine cytosine adenine) codes for the amino acid threonine Near universality (slight differences in mitochondrial DNA) Evolutionary significance Chapter 17 The Genetic Code: A Tool Chapter 17 Some things to be aware of… Although mRNA doesn’t necessarily start with AUG (start codon), nothing gets translated until the ribosome hits that point. Similarly, nothing gets translated after UAA, UGA, or UAG (stop codons). 3 Types of RNA Chapter 17 1. mRNA: The RNA transcript that exits the nucleus and is used by the ribosome to translate a polypeptide. 2. tRNA: The RNA located in the cytoplasm that brings amino acids to the ribosome to be added to a growing polypeptide. 3. rRNA: The RNA located in the ribosome that helps the ribosome to “read” the mRNA transcript. Chromosome Regions Chapter 17 1. Heterochromatin - highly condensed chromatin; areas that are not transcribed. 2. Euchromatin - less condensed chromatin; areas of active transcription. Chapter 17 Translation Overview • As mRNA goes through ribosome, its codons are matched up with anticodons on tRNA. • tRNA carries amino acids that correspond with the codons tRNA binds to in mRNA. • Amino acids are therefore attached in a specific order during translation. Transfer RNA (tRNA) 20 types of tRNA synthetases join 20 different amino acids to the correct tRNAs. There are ~45 different tRNAs. Wobble position allows some tRNAs to bind to more than one codon. Chapter 17 Wobble in tRNA Chapter 17 Due to evolution of the genetic code, the third base can be less discriminatory for the amino acid than the other two bases This third position in the codon is referred to as the wobble position. At this position Us and Cs may be read by a G in the anticodon. Polyribosomes A single mRNA can make many polypeptides simultaneously because several ribosomes can translate it at the same time Chapter 17 Chapter 17 Post Translation polypeptide chains fold into their functional tertiary structure tertiary proteins may join with other tertiary proteins becoming the subunits of a protein with quaternary structure some amino acids may be chemically modified enzymes may remove amino acids from the leading end or cleave a polypeptide chain into two pieces Rough ER Ribosomes Chapter 17 translation always begins on ribosomes in the cytoplasm if the mRNA codes for a protein destined for cell secretion or the endomembrane system, the ribosome will attach to the rough ER the cue to do so comes from a signal peptide (~20 amino acid sequence) in the growing polypeptide chain that is recognized by signalrecognition particle (SRP) in the cytoplasm Chapter 17 Mutations: changes in DNA Point mutations – changes in a single base-pair substitution – replacement of one nucleotide & its partner with another pair of nucleotides can be “silent” – have no effect on the encoded protein are usually missense – change a single amino acid in the encoded protein can be nonsense – change a codon to a stop codon causing translation to terminate early (shortens encoded protein) Frameshift mutations insertion – addition of nucleotide pairs deletion – loss of nucleotide pairs when insertions & deletions are not a multiple of 3, they significantly alter the reading frame (change all codons downstream of the mutation) http://www.youtube.com/watch?v=eDbK0cxKKsk Chapter 17 Mutations: changes in DNA Point mutations vs. Frameshift mutations Point – a nucleotide is just wrong Frameshift – a nucleotide is added or deleted. ACG TCC Point Shift at the ‘G’ ACA TCC ACG TCC Frame shift, G deleted ACT CC Chapter 17 Chapter 17 Chapter 17 Chapter 17 Mutagens physical & chemical agents that react with DNA in ways that cause mutations examples: X-rays, UV rays, carcinogens MUTAGENESIS- A process by which the genetic information of an organism is changed in a stable manner, either in nature or experimentally by the use of chemicals or radiation.