DNA Structure & Function (Outline)1. Historical perspective (DNA as the genetic material): Genetic transformation and DNADNA is the genetic material in bacterial viruses (phage)The base-pairing ruleDNA structure2. Basis for polarity of SS DNA and anti-parallelcomplementary strands of DNA3. DNA replication models4. Mechanism of DNA replication: steps and molecularmachinery5. Replication and the end of linear chromosomesMolecular basis for aging6. Fidelity of DNA replication
By definition the geneticmaterial of must be replicatedDNA Replication direct the cell functions byproviding information forproduction of proteinsFlow of the genetic information(Gene Expression)
DNA as the Genetic MaterialTime-line1850’s1870-1890190220th centuryMendelMicrosocopy: Mitosis and MeiosisChromosome basis of inheritance(Thomas H. Morgan)Work with bacteria and viruses
DNA StructurePrior to the 1950s,DNA is a polymerof nucleotidesconsisting of: a nitrogenous base a sugar a phosphate group
1928 Fredrick Griffith ExperimentsConcept of transformation (using Bacteriathat cause pneumonia in mammals)1944 Avery, McCarty, and MacLeodThe transforming material is DNA“DNA is the genetic material”1952 Hershey and ChaseDNA is the genetic material in viruses thatinfect bacteria
Griffith- Phenomenon of Transformation, a change ingenotype (genetic makeup) by a foreign substance thatchanges the phenotype (observed properties) of the cell
History of DNAAvery, MacLeod, and McCarty, 1944- DNA is the transforming material(Can convert Type R bacteria into S)
A phage, is a virus that infects bacteria and ismade of DNA and protein. Alfred Hershey and Martha Chase- thegenetic material of the phage T2 is DNA.Hershey Chase /sites/0072437316/student view0/chapter14/animations.html#
Biochemical analysis of DNA:Base-pairing rule1947 Erwin Chargaff, analysis of DNA from differentspecies %A %T & %C %GHuman DNAA 30.9%T 29.4%C 19.9%G 19.8%Class TP question
Base-pairing in DNA
Structural Model of DNAMaurice Wilkins and Rosalind Franklin- X-raycrystallography: Polynucleotide Helix(a) Rosalind Franklin(b)Franklin’s X-ray diffractionPhotograph of DNA
Watson and Crick deduced that DNA was adouble-stranded helixThrough observations of the X-ray crystallographic images of DNA
Chemical Structure of DNA HHMIPolarity and antiparallel nature ofthe two DNAstrands (5’ and3’ ends)
Watson and Crick- Specificity of pairing is dictated by the structure ofthe basesExample
Three models for DNA replication:Conservative modelSemi-conservative modelDispersive model
Meselson-Stahl ExperimentBacteriacultured inmediumcontaining15NDNA samplecentrifugedafter 20 min(after firstreplication)Bacteriatransferred tomediumcontaining14NDNA samplecentrifugedafter 40 min(after secondreplication)Meselson-Stahl Experiment animationLessdenseMoredense
The Basic concept of DNA replicationEach strand of DNA act as a template forsynthesis of new complementary strands
Major Events in the History of EarthRole of RNA in DNA ReplicationCenozoicHumansLand plantsOrigin of solarsystem deukaryotes41Proterozoic- Archaeaneoneon23ProkaryotesAtmosphericoxygenRNA
Molecular Mechanism of DNA ReplicationCollective action of several macro-molecules: DNA Proteins (enzymes & others) RNA Ribo-protein (for linear chromosomes)Direction of replication of new strands; 5’-----3’How nucleotides are added in DNA replication (Activelink)
DNA polymerase adds deoxyribonucleotides in a 5’ to 3’direction, it adds nucleotides to the 3′ end of a growingstrand.The new strand always starts with the 5’ end, the templatestarts with the 3’ end.
Primase, an RNA polymerase, uses the DNAtemplate strand to polymerize a shortcomplementary RNA chain (RNA primer)Two different DNA polymerases both- cannot initiate the synthesis of a polynucleotide- can only add nucleotides to an existing 3′ end
Summary of DNA ReplicationSemi-conservativeInitiation: Origin of replicationPrimase and RNA primerTemplate strand vs. new strand5’ to 3’ directionDNA polymerase (III and I)Base-pairing rulesdNTPs: deoxy-ATP, deoxy-GTP, deoxy-CTP, deoxyTTPLeading and lagging strandsOkazaki fragmentsDNA ligaseBidirectionalFidelity of DNA replication is maintained by activity ofDNA polymerase and other proof-reading systems.
Origin of ReplicationDNA Replication Tutorial (active link)
Other proteins participate in DNA replicationincluding: Helicase, topoisomerase, singlestrand binding protein
Replication of long DNA molecules begins atmultiple origins of replication simultaneouslyand is bidirectional
Replicating the Ends of linear DNA MoleculesMechanism ofDNA replicationcausestelomeres to getshorter with eachround ofreplicationLast fragmentLagging strand 5′Previous fragmentRNA primer3′Primer removed butcannot be replacedwith DNA becauseno 3′ end availablefor DNA polymerase3′Removal of primers andreplacement with DNAwhere a 3′ end is available5′Second roundof replication5′New leading strand ancer/images.htmlNew leading strand 5′3′Further roundsof replicationShorter and shorterdaughter molecules
Current ConnectionstoDNA structure and replicationQ: Why are we mortal with a limited lifespan?A: Our cells have a limited life span (# of celldivisions)
Telomerase- an enzyme(riboprotein) that extends the3’ end of the DNA strand byadding a repeated sequenceof 6-nucleotides typicallyTTAGGG (100-1000 times)https://www.youtube.com/watch?v AJNoTmWsE0shttps://www.youtube.com/watch?v vtXrehpCPEE
Ends of linear chromosomes have special DNAsequences and are known as telomeresadded by an enzyme known as telomeraseafter DNA replication is completed
Life span of dividing cells Telomerase is active in sperm, eggs, stem cells (bonemarrow), and cancer cells but not in somatic tissuesMost cells lose 50-200 endmost bases after each celldivisionAfter about 50 divisions, shortened telomeres signal thecell to stop dividingFigure 2.3
Fidelity of DNA replication & maintaining DNAintegrityMaintained by:1. Proof-reading function of DNA polymerase2. DNA repair smatch repair-lg.movDNA damage and repair in generalhttp://www.youtube.com/watch?v y16w-CGAa0Y&feature relatedhttp://www.youtube.com/watch?v nPS2jBq1k48
Genetic Integrity and Diversity Need for maintaining genetic integrity isbalanced by having enough geneticvariability for natural selection to act on Few errors of DNA replication are notcorrected!
Genetic transformation and DNA DNA is the genetic material in bacterial viruses (phage) The base-pairing rule DNA structure. 2. Basis for polarity of SS DNA and anti-parallel complementary strands of DNA 3. DNA replication models 4. Mechanism of DNA replication: steps and molecular machinery
1 DNA Structure & Replication (Outline) Historical perspective (DNA as the genetic material): Genetic transformation DNA as the transforming agent DNA is the genetic material in bacterial viruses (phage) Historical perspective (Structure of DNA): Identifying ribose and deoxy ribose
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DNA Structure and Replication 3 Model 2 - DNA Replication Direction of DNA helicase DNA helicase Free Nucleotides 11. Examine Model 2. Number the steps below in order to describe the replication of DNA in a cell. _ Hydrogen bonds between nucleotides form. _ Hydrogen bonds between nucleotides break. _ Strands of DNA separate.
Recombinant DNA Technology 3. Recombinant DNA Technology 600 DNA ISOLATION AND PURIFICATION Basic to all biotechnology research is the ability to manipulate DNA. First and foremost for recombinant DNA work, researchers need a method to isolate DNA from different organisms. Isolating DNA from bacteria is the easiest procedure because bacterial cells
2. At the end of DNA replication, (four/two) new strands of DNA have been produced, giving a total of (four/six) strands of DNA. 3. New DNA is replicated in strands complementary to old DNA because production of new DNA follows the rules of (base pairing/the double helix). Identifying Structures On the lines corresponding to the numbers on the .
The Insider’s Guide to DNA 1 Family history is in our DNA We all have DNA. It’s the genetic code that tells your body how to build you. You inherit half of your DNA from each parent: 50% from Mom and 50% from Dad, though exactly which DNA gets passed down is random. Because they inherited their DNA in the same way from their parents, your .
DNA cytosine methylation is a major epigenetic mark in eukaryotes. In plants, the DNA methyla-tion level in the genome is controlled by de novo DNA methylation, maintenance DNA methylation and DNA demethylation. De novo methylation is mediated by RNA-directed DNA methylation (RdDM), which can occur at all cytosine contexts,
On “Day & Date” watches, the days of the week are in English French. Once set in English, the consecutive days will continue to be in English. 3 OPERATING INSTRUCTIONS TO FIND THE INSTRUCTIONS THAT APPLY, SIMPLY MATCH YOUR WATCH TO THE DIAGRAMS ON THE FOLLOWING PAGES. SIMPLE TIME / MINI SWEEP To set the time: 1. PULL out crown to B .