Packet flashcards
Chromosomes
Complex of DNA and proteins
Genes
Made of DNA and act as instructions to make proteins
2 major features of DNA
The backbone made of sugar and phosphate groups.
Series of bases that project from each sugar in the backbone
Hershey-Chase Experiment
Studied how the T2 virus infects and replicates in the bacterium Escherichia coli
T2 infection of E. coli begins when:
The virus injects its genes into the cell and they direct the production of new virus particles.
What was the conclusion of the Hershey-Chase experiment
DNA can replicate
Structure of DNA
Double-stranded
Each strand consists of deoxyribonucleotides (deoxyyribose sugar, phasphate group, and nitrogenous base)
Phosphodiester linkage
Covalent bond
The hydroxyl group on 3’ carbon of one deoxyribose joined by a covalent bond to the phosphate group attached to 5’ carbon of another deoxyribose.
DNA directionality
5’ to 3’ direction
Semiconservative replication
Parental strands separate and each is template for a new daughter strand.
Each daughter has one old and new strand.
Conservative replication
The parental molecule serves as an entirely new molecule.
One daughter has both old strands; other has both new strands.
Dispersive replication
Parent molecule is cut into small pieces
Each daughter has an old and new DNA interspersed.
What did Watson and Crick propose
Proposed existing DNA strands of DNA served as a template.
Deoxyribonucleotides were added to new strands according to complementary base pairings.
Meselson-Stahl Experiment
Experiment done by Mehselson and Stahl that demonstrated that DNA replicated semi-conservatively.
Grew bacteria in a heavy isotope of Nitrogen (15N), then transferred to light nitrogen. Put in medium and spun.
DNA Polymersase
Enzyme that catalyzes DNA synthesis
What direction does DNA synthesis proceed
It only works in one direction which means the 5’ –> 3’ direction
Origin of replication
Sequence of bases on a chromosome where DNA replication starts
How many origin of replications are in bacterial chromosomes
1
How many origins of replications are there in eukaryotes
Multiple orgins of replication along each chromosome
What forms as DNA is synthesized
Replication bubble
Where do replication bubbles form
specific sequence of bases called the origin of replication.
Where does active DNA synthesis take place?
Replication forks of each replication bubble
DNA helicase
Protein that breaks hydrogen bonds between two DNA strands to separate them
Single-strand DNA-binding proteins
Attach to separating strands of DNA during replication. It prevents them from reforming the double helix
Topoisomerase
An enzyme that unwinds DNA double helix
3 limitations of DNA polymerases
- Can only synthesize DNA in the 5’–> 3’ direction
- DNA polymerases cannot start synthesis from scratch on a template strand
- DNA polymerases can only extend from the 3’ end of an existing strand that is hydrogen-bonded by complementary base pairings to the template
Primer
a short nucleic acid sequence that provides a starting point for DNA synthesis
Primase
An enzyme that synthesizes a short stretch of RNA to use as a primer during DNA replication
RNA polymerase
Enzyme that synthesizes RNA molecules from a DNA template through transcription
Difference between RNA and DNA polymerase
RNA polymerase can start synthesis from scratch
Leading strand
Strand of DNA that is synthesized towards the replication fork
Lagging strand
Strand synthesized away from the replication fork
Discontinuous replication hypothesis
Proposed to explain how the lagging strand is synthesized.
Held that primase synthesizes new RNA primers for lagging strands, and that DNA polymerase synthesizes short DNA fragments from these primers
Ozaki fragments
Short segments of DNA produced during replication of lagging strand template
Okazaki fragments are eventually linked together to produce the lagging strand in newly synthesized DNA
Synthesis of lagging strand
Priming: The lagging strand is synthesized in the 3’ to 5’ direction, which is opposite to the direction of the DNA replication fork. To initiate synthesis, an RNA primer is synthesized by the enzyme primase.
Initiation of Okazaki Fragment: DNA polymerase III adds nucleotides to the RNA primer, synthesizing a short DNA fragment called an Okazaki fragment. This process is initiated at the RNA primer.
Extension of Okazaki Fragment: DNA polymerase III continues to add nucleotides, extending the Okazaki fragment in the 5’ to 3’ direction.
Removal of RNA Primer: The RNA primer of each Okazaki fragment is removed by the enzyme RNase H, leaving a gap.
Fill-in by DNA Polymerase I: The gap left after primer removal is filled in by DNA polymerase I. This enzyme has both 5’ to 3’ polymerase activity and 5’ to 3’ exonuclease activity.
Ligation: DNA ligase seals the nick between adjacent Okazaki fragments by catalyzing the formation of a phosphodiester bond, producing a continuous lagging strand.
DNA ligase
An enzyme that joins pieces of DNA by catalyzing the formation of a phosphodiester linkage between the pieces.
DNA polymerase III
Extends leading strand and creates okazaki fragments by extension of RNA primers
Sliding clamp
Holds DNA polymerase in place during strand extension
Replisome
macromolecular machine that copies DNA; includes DNA polymerase, helicase, primase, and other enzymes
Telomeres
Region at the end of the eukaroyitc chromosome
Problem with copying telomeres
The lagging strands become too short as an enzyme that degrades the ribonucleotides removes the primer. As a result the single-strand DNA remains single stranded