Ch. 9: Molecular Biology Flashcards
Central dogma
DNA –> RNA –> protein –> trait
Griffith’s experiment
genetic info. can be transferred from dead bacteria to living bacteria (bacteria can uptake genetic info. from environment)
nonvirulent strain of pneumonia transformed into virulent strain when mixed w/ heat killed virulent strain
Transformation
ability of bacteria to absorb and express genetic information (DNA) obtained from their surroundings
Avery’s experiment
identified DNA as the heredity information of a cell
after removing protein coats from dead virulent bacteria stuff that was left was still able to transform bacteria (DNA)
Hershey and Chase experiment
est. that DNA was genetic material of phages
used simple structured phages (viruses that infect bacteria) injected w/ radioactive sulfur in proteins and radioactive phosphorus in DNA to show that DNA went into bacteria and it was DNA and not protein
Franklin, Watson, and Crick experiments
determine structure of DNA
x-ray diffraction photographs by Franklin helped Watson and Crick’s double helix twisted ladder model
One-gene-one-enzyme (polypeptide)-hypothesis
gene is defined as the segment of DNA that codes for a particular enzyme/ polypeptide
DNA replication (theory)
during interphase, a second chromatid copy of DNA is assembled
DNA molecule is unzipped, each strand serves as template to new, complementary strand. result is two identical double-stranded molecules of DNA
Semiconservative model
each new double-stranded molecule of DNA consists of template strand (old strand) and complementary strand (new, replicated DNA)
Helicase/ Replication fork
unwinds DNA helix during replication, forming Y-shaped replication fork
Single-stranded binding proteins
attach to each strand of uncoiled DNA during DNA replication to keep them separate
Topoisomerase
removes twists and knots that form in the double-stranded template as a result of the unwinding induced by helicase
DNA polymerase
enzyme that assembles the new DNA strand
moves in the 3’–>5’ so complement (new) strand is made in antiparallel, 5’–3’ direction
Leading strand
for the 3’–>5’ template strand, replication occurs continuously as DNA polymerase follows the replication fork, assembling a 5’–>3’ complementary strand
Lagging strand/ Okazaki fragments
DNA polymerase moves away from replication fork bc can only add nucleotides to the 3’ end
as DNA is uncoiled, DNA polymerase creates Okazaki fragments, requiring more time
DNA ligase
connects Okazaki fragments to produce a single complementary strand
Primase/ RNA primer
begins replication by forming RNA primer
leading strand and every Okazaki fragment on lagging strand must begin w/ RNA primer
DNA polymerase attaches to primer and makes DNA nucleotides
DNA replication (steps)
- Helicase unwinds, SSBPs and Topoisomerase keep apart
- Primase makes RNA primer to start
- DNA polymerase begins elongation
- Leading complementary strand assembled continuously
- Lagging strand assembled in Okazaki fragments
- Okazaki fragments joined by DNA ligase
- RNA primers replaced w/ DNA nucleotides
Prokaryotic vs. eukaryotic DNA replication
- Chromosome structure: prokaryotes have circular chromosome while eukaryotic chromosomes is linear w/ telomere ends
- Origins of replication: prokaryotic chromosome has one unique origin of replication while eukaryotes have multiple bc much larger chromosomes
Proofreading (DNA repair)
DNA polymerase checks if each newly added nucleotide correctly base-pairs with the template strand, if not the correct one is put in
Mismatch repair proteins (DNA repair)
repair errors that escape the proofreading ability of DNA polymerase
Excision repair proteins (DNA repair)
identify and remove damaged nucleotides caused by environmental factors like toxins or radiation
polymerase then uses the undamaged complementary strand as a template to repair damage
Excision repair proteins (DNA repair)
identify and remove damaged nucleotides caused by environmental factors like toxins or radiation
polymerase then uses the undamaged complementary strand as a template to repair damage
Protein synthesis steps
- Transcription: RNA molecules created by using one strand of DNA as template
- RNA processing: RNA is modified w/ deletions and additions
- Translation: processed RNA molecules used to assemble amino acids into a polypeptide
Replication of telomeres
during lagging strand replication RNA primer is removed and replaced w/ DNA nucleotides by DNA polymerase
problems that can occur when replication reaches end of DNA: not enough template strand remains for primase to attach/ at last primase no next Okazaki fragment for DNA polymerase to attach to so empty space is left
solution: enzyme telomerase attaches to end of template strand and extends template strand by adding repeat nucleotides… lagging strand will not lose DNA
Telomerase (aging)
telomerase active in young cells, but activity declines as cells age, eventually stopping; once telomerase stops, chromosome becomes shorter w/ each replication and DNA is slowly lost, resulting in non-viable daughter cells and aging affects seen
Kinds of RNA needed for protein synthesis (produced during transcription)
Messenger RNA (mRNA), Transfer RNA (tRNA), Ribosomal RNA (rRNA)
Messenger RNA (mRNA)
single strand of RNA that provides template (codons, 64 possible) for making amino acids into polypeptide
Transfer RNA (rRNA)/ Anticodon
short RNA molecule used for transporting amino acids to their proper place on the mRNA
nucleotides within tRNA interact/ pair and it folds into 3D molecule; one end of tRNA attaches to an amino acid, other end has the anticodon which binds w/ the codon on the mRNA during translation
Wobble pairing
exact base pairing between third nucleotide of the tRNA anticodon and the nucleotide of the rRNA codon often not required
allows the anticodon of some tRNAs to base pair w/ more than on kind of codon
Ribosomal RNA (rRNA)
combine w/ various proteins to form ribosomes
rRNA molecules are transcribed in the nucleolus and assembled w/ proteins imported from the cytoplasm to form a large and a small ribosome subunit… in cytoplasm these two subunits join to form ribosome (coordinates mRNA and tRNA during translation)
Transcription (steps)
- Initiation
- Elongation
- Termination
Initiation (transcription)
RNA polymerase attaches to promoter region (TATA box) and begins to unzip the DNA into two strands
Elongation (transcription)
occurs as RNA polymerase unzips DNA and assembles RNA nucleotides using one strand of the DNA as a template
5’ –> 3’
in contrast to DNA replication new nucleotides are RNA nucleotides and not DNA, only one DNA strand is transcribed and primers are not required
Termination (transcription)
RNA polymerase reaches special sequence of nucleotides that serve as termination point
5’ cap (mRNA processing)
guanine nucleotide w/ 2 additional phosphate groups, forming GTP
added to 5’ end of mRNA
provides stability to mRNA and point of attachment for small subunit of ribosome
Poly-A-tail (mRNA processing)
attached to 3’ end of mRNA
200 adenine nucleotides
provides stability to mRNA and controls movement of mRNA across nuclear envelope
RNA splicing (mRNA processing)
small nuclear ribonucleoproteins (snRNPs) delete introns (intervening non coding sequences) and splice exons (express polypeptide code)
Alternative splicing (mRNA processing)
selectively removing different parts of an RNA allows different mRNAs to be generated from the same RNA transcript
Where does translation occur?
cytoplasm
Translation (steps)
- mRNA attaches to ribosome
- sequence of codons on mRNA determines the sequence of amino acids in the polypeptide to be synthesized
- One by one, tRNA brings an amino acid to the ribosome such that the anticodon of the tRNA base-pairs w/ the codon of the mRNA
- newly arrived amino acid is attached w/ a peptide bond to the other amino acids already present
- tRNA released from ribosome
- process is repeated until “stop” codon on mRNA reached and full polypeptide made
Where does energy from translation come from?
GTP
Ribosome binding sites?
A, P and E
A site
for Amino acid/ Acceptor
in the first position, accepts an incoming tRNA carrying an amino acid to be then passed on to the tRNA in the second position
P site
for Polypeptide
in the second position, holds the tRNA w/ a growing chain of amino acids
E site
for Exit
in the third position, holds the tRNA after it gives up its amino acid
Initiation (translation)
- begins when small ribosomal subunit attaches near the end of the mRNA
- tRNA w/ anticodon UAC carrying methionine attaches to the mRNA at start codon AUG
- large ribosomal subunit attaches to the mRNA w/ the tRNA (that has methionine), occupying the P site (middle)
ribosome is now completely assembled w/ the mRNA and one tRNA
Elongation (translation)
- occurs as additional tRNAs arrive w/ their amino acids; newly arrived tRNA attaches to A site w/ anticodon of the mRNA codon
- amino acid on tRNA in P site is transferred to the amino acid on the newly arrived tRNA in the A site
- translocation occurs as the ribosome moves over one binding site, A site open for new tRNA; meanwhile, tRNA in E site is released, free to bind w/ its specific amino acid and provide another delivery to the mRNA
- Elongation as tRNA continues to bring amino acids, polypeptide growing one amino acid at a time
Termination (translation)
occurs when the ribosome encounters one of three STOP codons
completed polypeptide, last tRNA, and two ribosomal subunits are released… they can now attach to same/ another mRNA and repeat the process