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DNA molecule Gene 1 Gene 2 Gene 3 DNA template strand TRANSCRIPTION TRANSLATION mRNA Protein Codon Amino acid
DNA Signal Gene NUCLEUS Chromatin modification Chromatin Gene available for transcription Exon Intron Tail RNA Cap RNA processing Primary transcript mRNA in nucleus Transport to cytoplasm CYTOPLASM Transcription Gene Expression
mRNA in cytoplasm Translation CYTOPLASM Degradation of mRNA Protein processing Polypeptide Active protein Cellular function Transport to cellular destination Degradation of protein
Transcription
33 Promoter TATA box DNA strand 55 33 55 Transcription factors 55 55 33 33 RNA polymerase 55 55 55 33 33 RNA transcript Transcription initiation complex INITIATION
Promoter Transcription unit DNA Start point RNA polymerase 55 55 33 33
Promoter Transcription unit DNA Start point RNA polymerase 55 55 33 33 Initiation 33 33 1 RNA transcript 55 55 Unwound DNA Template strand of DNA
Promoter Transcription unit DNA Start point RNA polymerase 55 55 33 33 Initiation 33 33 1 RNA transcript 55 55 Unwound DNA Template strand of DNA 2 Elongation Rewound DNA 55 55 55 33 33 33 RNA transcript
Promoter Transcription unit DNA Start point RNA polymerase 55 55 33 33 Initiation 33 33 1 RNA transcript 55 55 Unwound DNA Template strand of DNA 2 Elongation Rewound DNA 55 55 55 33 33 33 RNA transcript 3 Termination 55 55 55 33 33 33 Completed RNA transcript
Elongation RNA polymerase Nontemplate strand of DNA RNA nucleotides 3 end Direction of transcription (“downstream”) Template strand of DNA Newly made RNA 33 55 55
Protein-coding segment Polyadenylation signal 33 3 UTR5 UTR 55 5 Cap Start codon Stop codon Poly-A tail G PPPAAUAAA AAA … NEXT: Add the Poly-AAAAAA tail and GTP Cap (UTR = untranslated region)
hn-RNA mRNA Coding segment Introns cut out and exons spliced together 5 Cap Exon1 Intron 55 Exon2 Intron 120 Exon 3 Poly-A tail 5 Cap 5 UTR3 UTR NEXT: (Eukaryotes) Remove Introns Also Alternative Splicing & Trans-splicing occurs SPLICING
RNA transcript (pre-mRNA) Exon 1Exon 2Intron Protein snRNA snRNPs Other proteins 55
RNA transcript (pre-mRNA) Exon 1Exon 2Intron Protein snRNA snRNPs Other proteins 55 55 Spliceosome
RNA transcript (pre-mRNA) Exon 1Exon 2Intron Protein snRNA snRNPs Other proteins 55 55 Spliceosome components Cut-out Intron (lariat) mRNA Exon 1 Exon 2 55
The RNA Tie Club was right!
RNA transcript (pre-mRNA) Exon 1Exon 2Intron Protein snRNA snRNPs Other proteins 55 55 Spliceosome snRNA ->
Amino acid attachment site (a) Two-dimensional structure Hydrogen bonds Anticodon 33 55 tRNA ->
Ribosome mRNA Signal peptide Signal- recognition particle (SRP) CYTOSOL Translocation complex SRP receptor protein ER LUMEN Signal peptide removed ER membrane Protein Insulin! SRP-RNA rRNA in ribosomes
GFP = Green Florescent Protein (from Jelly fish)
Enhancer TATA box Promoter Activators DNA Gene Distal control element Studying Promoters GFP
Enhancer TATA box Promoter Activators DNA Gene Distal control element Group of mediator proteins DNA-bending protein General transcription factors Studying Promoters GFP
Enhancer TATA box Promoter Activators DNA Gene Distal control element Group of mediator proteins DNA-bending protein General transcription factors RNA polymerase II RNA polymerase II Transcription initiation complex RNA synthesis GFP Studying Promoters
Control elements Enhancer Available activators Albumin gene (b) Lens cell Crystallin gene expressed Available activators LENS CELL NUCLEUS LIVER CELL NUCLEUS Crystallin gene Promoter (a) Liver cell Crystallin gene not expressed Albumin gene expressed Albumin gene not expressed GFP (blue) GFP (red) blue! red!
“Brainbow” GFP= Nobel Prize 2008 Martin Chalfie, Roger Tsien and Osamu Shimomura
TRANSCRIPTION RNA PROCESSING DNA RNA transcript 33 55 RNA polymerase Poly-A RNA transcript (pre-mRNA) Intron Exon NUCLEUS Aminoacyl-tRNA synthetase AMINO ACID ACTIVATION Amino acid tRNA CYTOPLASM Poly-A Growing polypeptide 33 Activated amino acid mRNA TRANSLATION Cap Ribosomal subunits Cap 55 E P A A Anticodon Ribosome Codon E