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Genetic Coding in Cells DNA • • • • Double stranded Deoxyribose sugar Bases: C,G A,T Self replicate vs. • • • • • RNA Single stranded Ribose sugar Bases: C,G,A,U Can’t self replicate mRNA, tRNA, rRNA Both contain a sugar, phosphate, and base. DNA Replication 1. DNA divides in half by splitting down the center of the ladder starting at one end. 2. Each rung separates in the middle, between the 2 bases. C splits from G and A from T. DNA Replication cont. 3. The cell has spare DNA Units. 4. The correct DNA unit attaches itself to the appropriate rungs on each of the 2 half-ladders as the DNA molecule splits. A spare AT, a spare TA, a spare GC, & a spare CG. 5. After the DNA finishes “unzipping” and the spare DNA units join up with the rungs on the half ladders, 2 identical molecules are formed. Introduction to The Central Dogma of Molecular Biology Protein Synthesis Flow of Information: DNA RNA Transcription Proteins Translation Transcription is the process by which a molecule of DNA is copied into a complementary strand of RNA. This is called messenger RNA (mRNA) because it acts as a messenger between DNA and the ribosomes where protein synthesis is carried out. Transcription occurs in the Nucleus. 1. RNA polymerase (an enzyme) attaches to DNA at a special sequence that serves as a “start signal”. 2. The DNA strands are separated and one strand serves as a template. 3. The RNA bases attach to the complementary DNA template, thus synthesizing mRNA. Transcription cont. 4. 5. 6. The RNA polymerase recognizes a termination site on the DNA molecule and releases the new pre - mRNA. Next the pre - mRNA under goes splicing. This is when the non-coding sequences or introns are eliminated. The coding mRNA sequence can be described as an exon. Now the mature mRNA leaves the nucleus and travels to the ribosome in the cytoplasm. Transcription Translation Translation is the process of decoding a mRNA molecule into a polypeptide chain or protein. Each combination of 3 nucleotides on mRNA is called a codon or 3-letter code word. Each codon specifies a particular amino acid that is to be placed in the polypeptide chain (protein). Translation •A three-letter code is used because there are 20 amino acids that are used to make proteins. A Codon OH P HO NH2 O N O N CH2 H P O O N O CH2 P NH N Guanine NH2 N O Arginine H O HO N O O HO Adenine N NH2 O N O CH2 N O OH H N N Adenine Translation •There is a total of 64 codons with mRNA, only 61 specify a particular amino acid. •There are more than 1 codon for each of the 20 amino acids. •The remaining three codons (UAA, UAG, & UGA) are stop codons, which signify the end of a polypeptide chain (protein). •Besides selecting the amino acid methionine, the codon AUG also serves as the “initiator” codon, which starts the synthesis of a protein. Translation Transfer RNA (tRNA) •Each tRNA molecule has 2 important sites of attachment. 1.anticodon, binds to the codon on the mRNA molecule. 2.The other site attaches to a particular amino acid. •During protein synthesis, the anticodon of a tRNA molecule base pairs with the appropriate mRNA codon. Met-tRNA Methionine 16 Pu 17 9 A 17:1 13 12 Py 10 1 2 3 4 5 6 U* 7 A C C 73 72 71 70 69 68 67 Py 59A* 66 65 64 63 62 C Pu 49 50 51 52 G T C y Py G* 22 23 Pu 25 G 26 2020:120:2A 27 1 28 29 30 31 Py* 47:16 47:15 43 44 42 45 41 46 47 40 47:1 39 38 Pu* U 34 U 35 C A 36 Anticodon Translation Ribosome: •Are made up of 2 subunits, a large one and a smaller one, each subunit contains ribosomal RNA (rRNA) & proteins. •Protein synthesis starts when the two subunits bind to mRNA. •The initiator codon AUG binds to the first anticodon of tRNA, signaling the start of a protein. Protein Synthesis: Translation Ribosome: •The anticodon of another tRNA binds to the next mRNA codon, bringing the 2nd amino acid to be placed in the protein. •As each anticodon & codon bind together a peptide bond forms between the two amino acids. Protein Synthesis: Translation Ribosome: •The protein chain continues to grow until a stop codon reaches the ribosome, which results in the release of the new protein and mRNA, completing the process of translation. Protein Synthesis: Translation Translation - Initiation fMet Large subunit E P A UAC 5’GAG...CU-AUG--UUC--CUU--AGU--GGU--AGA--GCU--GUA--UGA-AT GCA...TAAAAAA Small mRNA subunit 3’ Translation - Elongation Polypeptide Arg Met Phe Leu Ser Aminoacyl tRNA Gly Ribosome E P A CCA 5’GAG...CU-AUG--UUC--CUU--AGU--GGU--AGA--GCU--GUA--UGA-AT GCA...TAAAAAA mRNA 3’ Translation - Elongation Polypeptide Met Phe Leu Ser Gly Arg Aminoacyl tRNA Ribosome E P A CCA UCU 5’GAG...CU-AUG--UUC--CUU--AGU--GGU--AGA--GCU--GUA--UGA-AT GCA...TAAAAAA mRNA 3’ Protein Synthesis AMINE H H O N ACID C C ANYTHING R H Amino Acid Alanine OH H H Serine H O N C OH H C H H H H2O H H C H H C H O C C C C HO N H N H H C O N H O C C H H C H H HO H OH OH Translation - Elongation Polypeptide Met Phe Leu Ser Gly Arg Ribosome E P A CCA UCU 5’GAG...CU-AUG--UUC--CUU--AGU--GGU--AGA--GCU--GUA--UGA-AT GCA...TAAAAAA mRNA 3’ Translation - Elongation Polypeptide Met Phe Leu Ala Ser Gly Aminoacyl tRNA Arg Ribosome E P A CCA UCU 5’GAG...CU-AUG--UUC--CUU--AGU--GGU--AGA--GCU--GUA--UGA-AT GCA...TAAAAAA mRNA 3’ Translation - Elongation Polypeptide Met Phe Leu Ser Gly Arg Ribosome E Ala P A UCU CGA 5’GAG...CU-AUG--UUC--CUU--AGU--GGU--AGA--GCU--GUA--UGA-AT GCA...TAAAAAA mRNA 3’ Transcription And Translation In Prokaryotes 5’ 3’ 3’ 5’ RNA Pol. Ribosome mRNA 5’ Ribosome