•3. replication, repair, and dna rerere.pdf · dna replication in eucaryotes takes place only ......
TRANSCRIPT
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Molekulární biologie(KBC/MBIOG)
Ivo FrébortAlberts et al. (2008) Molecular Biology of the Cell, 5th ed.
Garland Science, New York
• 3. Replication, Repair, and Recombination of DNA
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Different rates of protein evolution – natural selection
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Semiconservative replication of DNA
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The chemistry of DNA synthesis
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DNA replication fork
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DNA polymerase
E. coli Pol I (928 aa, 109 kDa) monomer
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Editing by DNA Pol I
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Exonucleic proofreading by DNA polymerase
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RNA primer synthesis(DNA primase)
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The reaction catalyzed by DNA ligase
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DNA helicase – opening the replication fork
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Single-strand binding proteins straighten the unwound helix
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Sliding clamp prevents DNA polymerase from dissociation
E. coli Human Pol III PCNA protein
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Loading and unloading of DNA polymerase on the lagging strand
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Procaryotic replication fork
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Moving replication fork
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Moving replication fork, part II
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Moving replication fork, part III
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Strand directed mismatch repair in eucaryotes
MutS
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The „winding problem“ of the DNA replication
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DNA topoisomerase Ihelps rotation by reversible nicking of one strand
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DNA topoisomerase II makes transient double-strand break
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Single origin of DNA replication in procaryotes
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Methylation of the E. coli origin of replicationLag phase (10 min) – mismatch repairing system
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Initiation and completion of DNA replicationReplication bubble
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Mammalian DNA replication fork
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Eucaryotic chromosomes contain multiple origins of replication
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DNA replication in eucaryotes takes place only during the S-phase of cell cycle
Mammals 8h, yeast 40 min
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DNA replication in eucaryotes and the cell cycle
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Identification of replication origins (ARS) in yeast
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Large „Origin Recognition Complex“ initiates the replication
Yeast
Human
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Nucleosome assembly during replication
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Inheritance of histone modification
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Telomere replication
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Control of the lenght of telomeres
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DNA repair
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Spontaneous alterations requiring DNA repair
Red – oxidative damageBlue – hydrolysisGreen - methylation
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Depurination and deamination
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Pyrimidine base dimerization by UV light
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Formation of mutations during replication of damaged DNA
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Two major DNA repair pathways
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Recognition of unusual nucleotide by base-flipping(DNA glycosylases)
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Deamination of DNA nucleotides. Why not U in DNA?
Specific DNA glycosylase
T-G pairing glycosylase
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Repair of double strand breaks
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Non-homologous end-joining by Ku protein
„Quick and dirty“ repair
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General (homologous) recombination
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DNA hybridization, the principle of recombination
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Repair of single strand breaks by homologous recombinantion
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DNA synapsis catalyzed by RecA protein in E. coli
Rad 51 and Rad52 in humans
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Flawless repair of double strand breaks by homologous recombinantion
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Holliday Junction
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Crossovers by homologous recombination in meiosis
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Homologous recombination in meiosis
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Gene conversion by mismatch correction and proofreading
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Differences in general recombination in mitotic and meiotic cells
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Site-specific recombination
Some mobile genetic elements of bacteria –DNA-only transposons
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Cut-and-paste transposition
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The life cycle of a retrovirus
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Reverse transcriptase
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Transposition by a retrovirus (such as HIV) or a retroviral-like retrotransposon
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Transposition by a nonretroviral retrotransposon
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Conservative site-specific recombination
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Insertion of lambda DNA into bacterial chromosome
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The life cycle of bacteriophage lambda
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Conservative site-specific recombination can be used to turn genes on and off (mouse)