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The information of DNA is its nucleotide sequence DNA dictates the synthesis of proteins Proteins are the links between genotype and phenotype 1) Genotype what in the DNAGenes, allelic combination 2) Phenotype- physical expression of Genes. The EXPRESSION Gene expression process by which DNA directs protein synthesis 1) 2 stages: transcription and translation
Products of Gene Expression: One gene one enzyme: but not all proteins are enzymes (all enzymes are proteins though) Then: One gene one protein But many proteins are composed of several polypeptides, each with own gene Proteins are chain of amino acids Now: One gene- one polypeptide hypothesis
Basic Principles of Transcription and Translation RNA is the bridge b/w genes and the proteins for which they code Transcription is the synthesis of messenger RNA (mRNA) using info in DNA Translation is the synthesis of a polypeptide in ribosomes using information I the mRNA 1) Ribosomes- in Rough ER in cytoplasm -- The structure that synthesizes the proteins 2) Makes polypeptides in the ROUGH ER A primary transcript (pre-MRA) is the initial RNA transcript from anu genes prior to processing The central dogma is the concept hat cells are governed by cellular chain of command: DNA (transcribe) RNA (translate) Protein
Genetic Code 20 amino acids but 4 nucleotide bases in DNA 3 nucleotides correspond to the an amino acids
Codons (in RNA) : Triplets of Nucleotides Info from gene is based on a triplet code (in DNA) a series of non-overlapping, 3 nucleotide works The words of genes are transcribed into complementary non overlapping three nucleotides works of mRNA These words are then translated into a chain of amino acids, forming polypeptide 1) Turned into ribosomes then translated
2) TRANSLATION OCCURS IN RIBOSOMES INN CTYOPLASM DNA replicates into RNA and replaces the T with a U 1) Is a double check method to make sure that the nucleotides products are right During Transcription, one strand (template) provides the template for the RNA transcript 1) The template strand is always the same strand for a given gene 2) So is this one strand in all 46 chromosomes? Or one template per pair/chromosome? 3) * viruses have the ability to use 2 strands both for completely different things mRNA base triplets (codons) read in the 5 to 3 during translation 1) this is because its antiparallel to the DNA codons are read 5 to 3 each codon specifies the amino acids (one of 20), to be placed at the corresponding position along a polypeptide (a linear sequence)
Cracking the CODE - 4 bases 20 amino acids one of the 64 codons, 61 code for amino acids; 3 triplets are stop signals to end translation 1) could have multiple codons could code for the same amino acids genetic code is redundant but not ambiguous 1) no codon specifies more than one amino acid 2) there are multiple triplets for same amino acid Everytime I see CUU its LEUcine!! BUUUUUUUT it doesnt mean LEUcine will be only be seen as CUU. There are also UUA, and UUG and CUC, etc. (look at the chart slide 14) codons must be read in that correct reading frame in order for the specified polypeptide to be produced 1) the transcribed sequence is always in U2) dna is read 35 CAT
Transcription
Helicase, DNA POL (I and III), topimeraise , single binding protiens, RNA polymersase 1) RNA Polymerase Builds, synthesizes, RNA mRNA specifically separates DNA strands and builds complimentary RNA nucleotides (no helicase is needed anymore) as well as anything else one 1 form in rna polymerase in bacteria
at least 3 forms in eukary cellsrna polymerase II creates mRNA RNA synthesis- same base-pairing rules as DNA, except uracil substitutes thymine DNA sequences where RNA polymerase attaches = promoter In bacteria, the sequence signaling the end of transcription is called the terminator
Elongation rna polymerase untwists the double helix (same job as helicase) transcription makes about 40 nucletides (eurkaryotes) attach, read, create genes can be transcribed simultaneously several RNA polymerases
Eukaryotic Cells modify RNA after transcription RNA Processing= modification to pre-mRNA by enzymes prior to the exit into cytoplasm 1. Both ends of the primary transcript are usually altered 2. Some interior parts of the molecule are cut and others are spliced together
1) Alteration of mRNA Ends a. The 5 End receives a modified nucleotide 5 cap b. The 3 end gets a poly A- tail (adenine) c. Modifications share several functions i. Seem to facilitate the export of mRNA to the cytoplasm ii. Protect mRNA from hydrolytic enzymes iii. Help ribosomes attach to the 5 end (of mrna) Figure 17.10 RNA Processing = 4 parts (2 inside/ 2 outside) 5 end receives a modified a nucleotide 5 cap(GTP) The 3 end gets a poly- A tail Modifications share several functions 1) Seem to facilitate the export of mRNA to the cytoplasm 2) Protect mRNA from hydrolytic enzymes 3) Help ribosomes attach to the 5 end (site of protein synthesis)
Gene Splicing Most eukaryotic genes have long noncoding nucleotide stretches lying between coding regions 1) Noncoding regions= introns the portion of mRNA that never mean to be translated and will result in being cut out (stay IN the nucleus) Alternative splicing specific introns can be made into extrons cause they have what sequence we need create a whole new polypeptide 2) Exons= A bunch of cut segments that are bunched together, form an mRNA and then Exit the nucleus 3) RNA Splicing removes introns and joins exons Creates mRNA molecules with continuous coding sequence
Translation mRNA translated into protein with help of tRNA 1) tRNA (UPS guy) transfers the amino acids to the polypeptide are not Identical HYDROGEN BONDING produces a 3d molecule , roughly L shaped (The tRNA) Each is a single strand, is 80 nucleotides along Each carries a specific amino acid at one end Each has an anticodon on the other end; *located on tRNA* it is complementary to the codon of mRNA
Accurate translation has 2 steps: 1) Match b/w tRNA and coreesponding amino acid a. Accomplished by aminoacyl-tRNA synthetase b. One for each amino acid 2) Match b/w the tRNA anticodon and corresponding mRNA codon
Ribsomes
Ribosome couple tRNA anticodons with mRNA codons The two ribosomal subunits (large and small) are made of proteins and ribosomal RNA (rRNA) 3 sites: 1) A site accepting site- accept charged trna molecules into the ribosomes Amino acid 2) P site- holds tRNA that carries polypeptide chain Peptidyl 3) E site exit side, where discharged tRNA leaves Exit
Building a Polypeptide - 3 stages of translation 1) Initiation 2) Elongation 3) Termination - All three stages require protein factors that aid in the process Ribosome Association and Initiation of Translation First, small ribosomal subunits binds with mRNA and special initiator rTNA Small subunit moves along the mRNA until it finds AUG (Met) [ will find UAC ] Proteins called initiation factors bring in the large subunit that completes the translation initiation complex
Elongation of the Polypeptide Chain Amino acid are added to the preceding amino acid at C-terminus (carboxyl) of the growing polypeptide chain Amino group N- Terminus (nitrogen) Each addition involves elongation factors 1) Codon recognition 2) Peptide bond formation 3) Translocation Translation proceeds along the mRNA in a 5 to 3 direction
Termination of Translation Occurs when a stop codon reaches the A site of the ribosome The A site accepts a release factor (protein) Causes the addition of water molecule instead of an amino acid releasing the polypeptide Translation assembly then comes apart (disassociates)
Polyribosomes A number of ribosomes can translate a single mRNA simultaneously, forming a polyribosome Polyribosome enable a cell to make copies of polypeptide quickly