L11: Genome Replication and Expression Flashcards
Overview of DNA replication
bidirectional
semi-conservative
strands separate
each strand is a template for the synthesis of a new strand
What’s different about DNA replication in eukaryotes and prokaryotes?
Eukaryote- multiple oris
Prokaryote- single ori
How is the origin of replication selected in prokaryotes?
Protein DnaA binds and “opens” ori
What enzyme catalyzes DNA synthesis in 5’ to 3’ direction?
DNA polymerase
What does DNA polymerase need for DNA synthesis?
template
deoxynucleotide triphosphates
primer (usually RNA with 3’ OH group)
What enzyme breaks H bonds between DNA strands?
helicase
What enzyme underwinds DNA and relieves supercoiling?
Gyrase (topoisomerase)
What enzyme lays down RNA primers?
DNA primase
What enzyme removes RNA primers and fills in the gaps?
DNA pol I
What enzyme is a target for ciprofloxacin antibiotic?
Gyrase
How are genetic instructions stored and organized in DNA?
DNA sequence of a gene corresponds to amino acid sequence of protein encoded
What enzyme is used for transcription in bacteria?
RNA polymerase
What is the target of rifampin (antibiotic)
RNA polymerase
What are sigma factors?
proteins that direct core of RNA polymerase to promoters
What is sigma^s important for?
stationary phase
What are the 3 types of RNA generated by transcription in bacteria?
tRNA
rRNA
mRNA
What is tRNA used for?
carries amino acids during protein synthesis
What is rRNA used for?
component of ribosomes
What is mRNA used for?
template for protein synthesis
What are the key features of the structure of RNA (that make it different from DNA)
ribose instead of deoxyribose
uracil replaces thymine
usually single-stranded
What is the terminator in transcription?
sequence that signals RNA pol to stop
What enzyme opens and unwinds DNA without the need for a primer?
RNA polymerase
When does the sigma factor leave?
after transcription is initiated
What are the two types of terminators?
Rho-independent transcription termination
Rho-dependent transcription termination
What type of nucleic acid is a terminator?
DNA
Which type of terminator causes RNA pol to release?
Rho-independent
What type of terminator causes RNA pol to pause (initially)
Rho-dependent
What does the protein Rho do to stop transcription?
binds to RNA, then moves toward RNA pol and causes release
How do bacteria control gene transcription in response to their environment?
two-component signal transduction systems
Where does transcription in prokaryotes occur?
cytoplasm
Where does transcription in eukaryotes occur?
nucleus
What do eukarya need for transcription?
RNA polymerase Transcription factors (no sigmas)
What is the promoter element for transcription in Eukarya?
TATA box
What process removes introns?
RNA splicing
What modifications need to be made to mRNA in Eukarya?
capping - methylguanosine added at 5’ end
Polyadenylation - adenine nucleotides added at 3’ end - tail
What is translation?
synthesis of polypeptides directed by mRNA sequence
What does translation require?
ribosomes
energy in the form of ATP and GTP
What are the 2 subunits of bacterial ribosomes?
30S and 50S
What type of rRNA acts as a ribozyme
23S rRNA
What type of rRNA is capable of peptidyltransferase
23S rRNA
What type of rRNA aligns mRNA with the ribosome?
16S rRNA
What type of rRNA has a sequence complementary to Shine-Dalgarno sequence of the mRNA
16S rRNA
What do ribosomes do?
“read” mRNA as a code
code is ____ - multiple codons can encode the same amino acid
degenerate
codons are read as a ____ made up of __ nucleotides
triplet, 3 nucleotides
How many codons are there?
64
How many nonsense codons are there?
3 (stop codons)
How many sense codons are there?
61 (specify amino acids)
What enzyme attaches an amino acid to the 3’ end of the correct tRNA molecule?
Synthetase
What is the shape of tRNAs?
clover
What is the anticodon?
complementary to codon in mRNA
What are the 3 stages of translation?
initiation
elongation
termination
What are the 3 sites on the ribosome?
A
P
E
What occurs during initiation of translation?
AUG codon binds to P site, tRNA attaches Met amino acid
What occurs during elongation of translation?
Next tRNA binds to A site, peptide bond forms between P and A site amino acids, everything shifts down and process continues
What occurs during elongation of translation?
Next tRNA binds to A site, peptide bond forms between P and A site amino acids, everything shifts down and process continues
What occurs during termination of translation?
Any one of the 3 stop codons enters, no tRNAs carry anticodon
release factors cleave and release the polypeptide
What binds to single-stranded DNA after strands are separated to keep them separated?
SSB proteins (single-stranded binding proteins)
What enzyme has 3’ -> 5’ proofreading activity?
DNA pol III