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From Gene to Protein. How Genes Work. What do genes code for?. How does DNA code for cells & bodies? how are cells and bodies made from the instructions in DNA. DNA. proteins. cells. bodies. The “Central Dogma”. Flow of genetic information in a cell
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From Gene to Protein How Genes Work
What do genes code for? • How does DNA code for cells & bodies? • how are cells and bodies made from the instructions in DNA DNA proteins cells bodies
The “Central Dogma” • Flow of genetic information in a cell • How do we move information from DNA to proteins? transcription translation RNA DNA protein trait replication
aa aa aa aa aa ribosome aa aa aa aa aa aa From gene to protein nucleus cytoplasm transcription translation DNA mRNA protein trait
Transcription fromDNA languagetoRNA language
RNA Review… • ribose sugar • N-bases • uracil instead of thymine • U : A • C : G • single stranded • Many types of RNA • mRNA, tRNA, rRNA, siRNA… transcription DNA RNA
Transcription • Making mRNA • transcribed DNA strand = template/coding strand • enzyme • RNA polymerase • RNA polymerase is versatile (it slices, it dices!) • Unwinds, unzips, adds in complementary RNA bp’s 3 A G C A T C G T 5 A G A A A C G T T T T C A T C G A C T DNA 3 C T G A A 5 T G G C A U C G U T C unwinding 3 G T A G C A rewinding mRNA template strand RNA polymerase 5 build RNA 53
Initiation: Where to begin? • Promoter region • = binding site before beginning of gene • “TATA box” binding site indicates location • “Upstream” of gene to transcribe/copy • = binding site for RNA polymerase
RNA polymerase Elongation A • Match RNA bases to DNA bases on one of the DNA strands • 5’ 3’ direction C U G A G G U C U U G C A C A U A G A C U A 5' 3' G C C A T G G T A C A G C T A G T C A T C G T A C C G T
Termination • Eventually… • RNA transcript is released • RNA polymerase detaches (complete mechanism still not fully known)
intron = noncoding (inbetween) sequence exon = coding (expressed) sequence Eukaryotic genes have junk! • Eukaryotic genes are not continuous • exons = the real gene • expressed / coding DNA • introns = the junk • inbetween sequence intronscome out! eukaryotic DNA
intron = noncoding (inbetween) sequence exon = coding (expressed) sequence mRNA splicing • = “Post-transcriptional processing” • eukaryotic mRNA needs work after transcription • primary transcript = pre-mRNA • mRNA splicing • edit out introns • make mature mRNA transcript ~10,000 bases eukaryotic DNA pre-mRNA primary mRNA transcript ~1,000 bases mature mRNA transcript spliced mRNA
Splicing must be accurate • No room for mistakes! • a single base added or lost throws off the reading frame AUGCGGCTATGGGUCCGAUAAGGGCCAU AUGCGGUCCGAUAAGGGCCAU AUG|CGG|UCC|GAU|AAG|GGC|CAU Met|Arg|Ser|Asp|Lys|Gly|His AUGCGGCTATGGGUCCGAUAAGGGCCAU AUGCGGGUCCGAUAAGGGCCAU AUG|CGG|GUC|CGA|UAA|GGG|CCA|U Met|Arg|Val|Arg|STOP|
snRNPs snRNA intron exon exon 5' 3' spliceosome 5' 3' lariat 5' 3' exon exon mature mRNA excised intron 5' 3' RNA splicing enzymes
Alternative splicing • Alternative mRNAs produced from same gene • when is an intron not an intron… • different segments treated as exons
3' poly-A tail 3' A A A A A mRNA 50-250 A’s 5' cap P P P 5' G More post-transcriptional processing • Need to protect mRNA on its trip from nucleus to cytoplasm • Enzymes in cytoplasm attack mRNA • protect the ends of the molecule from degrading • add 5 GTP cap • add poly-A tail • longer tail = mRNA lasts longer: produces more protein
aa aa aa aa aa ribosome aa aa aa aa aa aa From gene to protein nucleus cytoplasm transcription translation DNA mRNA protein trait