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RNA Transcription and Translation How is information in DNA used to make protein? RNA acts as ____________________ DNA and protein synthesis machinery Transcription - copying of ___________ __________________________ Translation - production of __________ __________________________ Whole process = _________________ DNA (gene) RNA Protein 1 RNA Ribonucleic Acid dAMP DNA uses _______________ RNA uses _______________ In RNA uracil is used instead __________________________ AMP UMP GMP CMP 2 RNA Ribonucleic Acid Formation of sugar-phosphate backbone As with DNA: - Individual nucleotides are connected through linking the ______________ ____________ with the phosphate group on the __________________________ Single strand of polynucleotides - RNA has directionality, 5’ - 3’ 5’ - A C G U - 3’ 5’ end = start of gene 3’ end = end of gene Ultimately -5’ end of RNA (gene) = _______________ -3’ end of RNA (gene) = _______________ Unlike DNA: RNA is ____________________, __________________ __________________ 3 RNA Ribonucleic Acid single stranded RNA can fold and _____ and _____ will H-bond forming base pairs H-bonding 4 RNA Ribonucleic Acid Six kinds of RNA transfer RNA (tRNA) - carries _____________________ during ribosomal RNA (rRNA) protein synthesis messenger RNA (mRNA) - is ___________________ - encodes ______________ small nuclear RNA (snRNA) micro RNA (miRNA) small interfering RNA (siRNA) 5 Transcription Central dogma of molecular biology DNA (gene) RNA Protein Transcription: the synthesis of ____________________________ 6 Transcription RNA polymerase – reads _________________________________ - ___________________ of DNA in 3’ - 5’ direction - resulting RNA is ________________________ - resulting RNA is same as ___________________ Transcription http://www.youtube.com/watch?v=Jqx4Y0OjWW4&feature=related 7 Transcription mRNA (messenger RNA) - the RNA produced from _____________ - is used for ____________________ - is capped at _______________ - is capped at ________________________ _______________________________ - both caps help ________________________ 5’ 3’ AAAAAAAAAAAAAA 8 Translation Central dogma of molecular biology DNA (gene) RNA Protein Transcription: the synthesis of RNA based on DNA sequence Translation: Synthesis of protein from RNA template Players in translation: - mRNA - ____________ - ____________ - ____________ 9 Translation Steps of translation: - _____________ - _____________ - _____________ How to read the sequence of mRNA? How does the sequence of mRNA translate into protein sequence? 10 Translation The genetic code Genetic code is: - _______________________________ code for one amino acid - _____________________ - continuous, ________________ __________________________ - universal, ___________________ ____________________ 11 Translation Codon - triplet of nucleotide bases _____________________________ Since 4 different nucleotide bases (A T (U) G C ) there are _______________________________ There are 20 different amino acids - ______________________________ for a single amino acid DNA sequence: ATG TTT TCT CCG AAT AGC GGT TGA transcription mRNA sequence: ___ ___ ___ ___ ___ ___ ___ ___ translation ___ ___ ___ ___ ___ ___ ___ ____ 12 Why multiple Codons for single amino acid? DNA sequence: ATG TTT TCT CC_ AAT AGC GGT TGA transcription mRNA sequence: AUG UUU UCU CC_ AAU AGC GGU UGA translation Met Phe Ser ___ Asn Ser Gly stop Mutation in DNA: ATG TTT TCA CC_ AAT AGC GGT TGA transcription mRNA sequence: Multiple codons______ ___________________ ___________________ ___________________ ___________________. AUG UUU UCA CC_ AAU AGC GGU UGA translation Met Phe Ser ___ Asn Ser Gly stop 13 Cracking the genetic code Marshall Warren Nirenberg (1927-2010) - American born biochemist - Worked at National Institutes of Health (NIH) - Genetic code experiments in early 1960’s. - Definitive experiment in 1964 - Nobel prize in physiology or medicine 1968 Har Gobind Khorana (1922-2011) - born in what is now Pakistan, biochemist - worked at Univ. Wisconsin - Madison - Nobel prize in physiology or medicine 1968 with Nirenberg 14 Translation How to convert mRNA sequence into protein? Process requires many different proteins and other factors. Two main players: Ribosome - a cellular structure that catalyses the __________________ _______________________________________ transferRNA (tRNA) - small RNA which binds to and carries single amino acids to __________________________ 15 Translation 1. Initiation: Ribosome binds to mRNA and moves along until it recognizes _____________________________________ 2. tRNA with bound amino acid binds to mRNA codon. - anticodon: - __________________________ - the complement ____________ 3. Elongation: Next tRNA binds to _______________________________ 4. Ribosome moves on to next codon, process repeats Met Phe Ser Pro Asn Ser 5. Termination: polypeptide chain is terminated when ____________________________ ______________________________ Met 5’ Phe Ser AAA AGA 3’ XXX AUG UUU UCU CCG AAU AGC GGU UGA AAAAAAAAAAAAAA 16 Gly tRNA Discovery Robert W. Holley (1922-1993) - American born biochemist - worked at Cornell University - finalized the structure of tRNA in 1964 - awarded Nobel prize in physiology or medicine 1968 along with Nirenberg and Khorana Translation movie: http://www.youtube.com/watch?v=5bLEDd-PSTQ&feature=related 17 tRNA Discovery Robert W. Holley - American born biochemist - worked at Cornell University - finalized the structure of tRNA in 1964 - awarded Nobel prize in physiology or medicine 1968 along with Nirenberg and Khorana Translation movie: http://www.youtube.com/watch?v=5bLEDd-PSTQ&feature=related 18 The Big Picture - what have we studied so far? Historically, this is the path of determining the ______________ _______________ How to go from DNA base pair sequence to protein amino acids sequence. Proteins: - 1˚, 2˚, 3˚, 4˚ structure - fibrous proteins - globular proteins - purification/analysis techniques - enzyme kinetics - mechanisms of enzyme control DNA: - structure - replication Transcription: - RNA polymerase - mRNA production Translation: - Ribosome - tRNA - a.a.’s linked to make protein Amino acids: - structure - acid/base properties 19 - peptide bond