Exam 1- Ch 10-14 Flashcards
Characteristics of Genetic Material
- Genetic material must contain complex info
- Genetic material must replicate faithfully
- Genetic instructions must encode the phenotype
- Must have capacity to vary
- Genetic material must contain complex info
- Store large amounts of info
- Instructions for traits and functions of an organism
- Genetic material must replicate faithfully
- Begins w single cell-> undergoes billions of divisions
- W/ each division, genetic instructions must be accurately transmitted to decendant cells
- When organisms reproduce and pass genes to progeny, genetic instructions must be copied w/ fidelity
- Genetic instructions must encode phenotype
- Genotype must have capacity to be expressed as a phenotype (code 4 traits)
- Product of gene= protein or RNA molecule
- -Must be mechanism 4 genetic instructions in DNA to be copied to RNAs and proteins
- Must have capacity to vary–> Genetic info must have ability to vary
-Diff species and individual members of species differ in genetic makeup
Chargaff’s Rule
- Developed by Chargaff and colleges concerning ratios of bases in DNA
- DNA from diff organisms varies in base comp
- A=T, G=C
Transforming principle
- Substance responsible 4 transformation
- Ex: DNA
Griffith Experiment
- Virulent strains= disease causing (S)
- Nonvirulent strains= non-disease causing (R)
- R strains lived, S strains killed, heat killed R mice lived, heat killed S w/ R died
- Conclusion= virulence is heritable
Avery, Macleod, and McCarthy experiment
- Showed DNA= transforming principle
- Virulent bacteria heat killed and added to nonvirulent
- Treated w/ RNAase, Protease, and DNAase
- RNAse and Protease–> virulent and nonvirulent bacteria
- DNAase–> only virulent bacteria
Hershey-Chase experiemnt
-T2 bacteriophage= virus that infects ecoli, reproduces by attaching to outer wall of cell and injecting its DNA into cell, cell synethsizes phage proteins
Q?- does phage protein or DNA transmitted in phage production
-DNA contains phosphorus, tagged w/ P isotope
-Protein contains Sulfer, tagged w S isotope
-S batch had no radioactivity, protein not transmitted
-P batch was radioactive, indicating DNA transmitted
Deoxyribose
- 5 carbon sugar in DNA
- Lacks a hydroxyl group on 2’ carbon atom
- Hydroxyl group on 1’ and 3’
Nitrogenous base
- Nitrogen-containing base
- one of the 3 parts of a nucleotide
Purine
- Double-ringed
- Type of nitrogenous base in DNA and RNA
- Adenine and Guanine
Pyramidine
- Single ringed
- Nitrogenous bases in DNA and RNA
- Cytosine, Thymine (not in RNA), Uracil ( not in DNA)
Phosphate Group
- A phosphorus atom attached to 4 oxygen atoms
- One of 3 compoents of nucleotide
- 5’ end
Deoxyribonucleotides
- Basic building block of DNA
- Consisting of deoxyribose, phosphate group, and nitrogenous base
Ribonucleotides
- Basic building block of RNA, consisting of ribose, a phosphate group, and a nitrogenous base
- Ribose has OH on 1’, 2’ and 3’
Phosphodiether linkages
- A strong covalent bond that joins the 5’ phosphate group of one nucleotide to the 3’ hydroxyl group of the next nucleotide in a polynucleotide strand
- Connection of nucleotides
3’ end
- End of a polynucleotide chain
- OH group is attached to 3’ carbon of sugar in nucleotide
B-DNA
- Right-handed helical strucutres of DNA
- exist when water is abundent
Primary Structure
- DNA as nucleotide structure
- How nucleotides join together
Secondary Structure
- 3D congifuration
- Helical structure
- H bonds link base pairs (3 bonds btwm G and C, 2 btwn T and A)
- DNA strands= antiparallel
DNA/ RNA backbone
-Alternating sugars and phosphate groups
Supercoiling
- Tertiary structure
- Forms when strain is placed on DNA helix by over or underwinding
- Takes up less space than relaxed DNA
Relaxed state
-Energy state of DNA molecule where there is no structural strain on molecule
Positive Supercoiling
- Tertiary structure
- Forms when strain is place on DNA helix by overwinding
Negative Supercoiling
- Tertiary structure
- Forms when strain is placed on DNA helix by underwinding
- Most DNA= neg supercoiled
- Advantages= separation of 2 strands easier during replication/ transcription, packed into smaller spaces
Topoisomerases
- Enzymes that add or remove rotations in a DNA helix by temporarily breaking nucleotide strands, rotating ends around each otherm, and rejoining broken ends
- Induces and relieves supercoiling
Euchromatin
- Chromatin that undergoes condensation and decondensation in course of cell cycle
- Majority of chromosomal material, where transcription takes place
Heterochromatin
- Chromatin that remains in highly condensed state throughout cell cycle
- Found at centromeres and telomeres
- Y chromosome= largely heterochromatin
- Lack of transcription
- Absence of crossing over and replication in S phase
Bacterial Chromosome
- Circular
- Looks like a clump
- Additional DNA in form of plasmids
Eukarytoic Chromosomes
-1 chromosome= 1 linear molecule of DNA (packed and folded)
Chromatin
- Complex of DNA and proteins
- Types= Euchromatin and Heterochromatin
- When in condensed form, nucleosomes fold on themselves into 30nm loop fibers, each anchored to base by proteins
Histones
- Small, + charged proteins
- H1, H2A, H2B, H3, H4
- Charge attracts DNA
Nucleosome
- Core of protein and DNA produced by digestion w/ nuclease enzymes
- Simplest level of chromatin structure (basic repeating unit)
- 146 bp of DNA wrapped 2 times around 8 histone (2 copies of H2A, H2B, H3, H4)
- H1- Binds bp of DNA where DNA joins and leaves histone octamer and locks DNA into place
- Separated by linear DNA
DNAase I
-Enzyme that digests DNA
Semiconservative Replication
- Replication in which 2 nucleotide strands of DNA separate and each serveds as a temple for synthesis of a new strand
- All DNA replication is semiconservative
Replicon
-Unit of replication consisting of DNA from the origin of replication to the point at which replication on either side of the origin ends
Origin of Replication
-Site where DNA synthesis is initiated
Theta Replication
- Replication of circular DNA (Bacteria)
1. Double stranded DNA unwinds at origin of replication, producing ingle nucleotide strands to serve as templates 4 new DNA
2. Unwinding of double helix creates replication bubble
3. DNA replication on both template strands= simultaneous w/ unwinding (2 forks= bidirectional) - Produces 1 new and 1 old strand
Replication Bubble
-Segment of DNA molecule that is unwinding and undergoes replication
Replication fork
-Point at which a double-stranded DNA molecule separates into two single strands that serve as templates for replication
Bidirectional Replication
-Replication at both ends of replication bubble
Rolling Circle Replication
- Replication of Circular DNA (Viruses)
1. Initiated by break in one of the nucleotide strands, exposing 3’ OH and 5’ Phosphate
2. New nucleotides are added to 3’ end of broken strand using inner (unbroken) strand as template
3. New nucs. added to 3’, 5’ displaced from template
4. 3’ grows around circle w each revolution around circle, 3’ displaces nuc strand in preceeding rev
5. Linear DNA molecule cleaved from O - *Result= Double stranded O dna molecule and single stranded dna molecule
DNA Polymerase
-Enzyme that synthesizes DNA
Continuous Replication
- Replication of leading strand of DNA in same direction as unwinding
- Allows new nucleotides to be added continuously to the 3’ ends of the new strands as template is exposed
Discontinous Replication
- Replication of the lagging strand of DNA in direction opposite to unwinding
- DNA must be synthesized in short fragments (Okazaki fragments)
- -Frags eventually joined together
Conservative replication
- False
- Entire double-stranded DNA molecule serves as template 4 replication
Dispersive Replication
- Both nucleotide strands break down into fragments and reassemble into new DNA strands
- New molecules contain old and new frags
- None of original molecule is conserved
Meselson and Stahl Experiment
-Used nitrogen isotopes of different weights and a centrifuge to prove that DNA is semiconservative in replication
Linear Eukaryotic Replication
- Multiple origins of replication
- A each origin, DNA unwinds and produces replication bubble
- Replication takes place at both strands at each end of the bubble w/ 2 replication forks spreading outward
- Replication forks run into each other, fuse to form large stretches of DNA
Requirements of Replication
- Template consisting of single-stranded DNA
- Raw materials to be assembled into new nucleotide strand
- Enzymes and other proteins that read template and assemble DNA molecule
DNA Replication
5’–> 3’ direction, nucs added to 3’ end
Initiator Proteins
- Proteins that bind to an origin of replication
- Unwinds a short stretch of DNA, allowing helicase and other single-strand-binding proteins to bind and initiate replication
DNA Helicase
- Enzyme that unwinds double-stranded DNA by breaking hydrogen bonds
- cannot initiate strands unwinding
- Binds to laggin strands temp at replication fork and moves in 5–>3 direction along strands (moves rep fork)
Single-Stranded-Binding Proteins (SSBs)
- Protein that binds to single-stranded DNA during replication after helicase
- Protect single-strand nuc chains and prevent seconardy structures
- Indiff to base sequence–> bind to any single-stranded DNA
- Form tetramer
DNA Gyrase
- Topoisomerase enzyme in ecoli that relieves torsional strain that builds ahead of replication fork
- Creates double-strand breaks
- Inhibition results in cessation of DNA synthesis
Primase
- Enzyme that synthesizes short stretch of RNA on DNA template
- Fxns in replication to provide a 3’ OH group for attachment of DNA nucleotide
Primers
- Short stretch of RNA on a DNA template
- Provides a 3’ OH group for the attachment of DNA nucleotide at initiation of replication
DNA Polymerase III
- Bacterial DNA polymerase that removes RNA primers and places them w DNA nucleotides
- Main replicator, adds nucs to 3’ end (5–>3 polymerase activity
- 3’–>5’ exonuclease acitivity (removes nucs in this direction to remove errors)
- High processivity
B Sliding Clamp
- A ring-shaped polypeptide component of DNA pol III,
- Encircles DNA during replication and allows pol to slide along DNA template strand
DNA Pol I
- Bacterial DNA pol that removes RNA primers and replaces them w DNA nucleotides
- Has 5–3’ polymerase, exonuclease and 3’–>5’ exonuclease activity
- –Removes primers and replaces w/ DNA nucs after DNA pol III initiated sythesis at downstream primers
Proof-reading
-Process by which DNA pols remove and replace incorrectly paired nucleotides in the course of replication
Mismatch Repair
- Process that corrects mismatched nucleotides in DNA after replication has been completed
- Enzymes excise take out nuc and use original nuc strand to replace it
Initiation, Replication - Bacteria
- 1 origin of replication
- Initiator protein binds to origin and causes DNA to unwind (allows helicase to attach to strand)
Unwinding, Replication-Bacteria
- DNA helicase
- SSBs
- DNA Gyrase
Elongation, Replication -Bacteria
- All DNA pol require nuc w/ 3’ OH group to add nucs to
- All newly synthesized DNA molecules have short RNA primers embedded in them, later removed and replaced w/ DNA nucs
- Lagging strand= new primer generated at beginning of each Okazaki
- Leading strand= primer required at 5’ end of new strand
- Primase forms complex w helicase at fork
- DNA Pol I and III
DNA Ligase
- Break that remains in sugar-phosphate backbone of new DNA strand is sealed
- 3’ OH attaches to 5’ phosphate group
Termination–Bacteria
-Terminates when replication forks meet or reach termination sequences
Origin-Recognition Complex
- Eukaryotes
- Multiprotein complex that binds to an origin of replication
- Unwinds the DNA around it to initiate replication
Replication Licensing Factors
- Eukaryotes
- Protein that ensureds replication takes place only once at each origin of replication
- Required at origin b4 replication can be initiated and removed after DNA is replicated
- Ensures DNA is not replicated until cell has passed through mitosis
DNA Polymerase a (alpha)
- Eukaryotes
- DNA pol that initiates replication by synthesizing RNA primer, followed by short string of DNA nucleotides
- Primase activity
- High fidelity
DNA pol d (delta)
- Eukaryotic
- DNA pol that replicates lagging strand during DNA synthesis
- Carries out DNA repair and translesion DNA synthesis
- High fedelity