Microbial Genetics Flashcards
(43 cards)
Where are microbial genomes located?
Eukaryotes - nucleus
Prokaryotes - nucleoid region & extrachromosomal plasmids
Viruses - inside the (nucleo)capsid
Plasmids
Small circles of dsDNA
Encode inessential genes that give the bacterium a selective advantage
Antibiotic resistance
Pili (conjugation) F- –> F+
The Central Dogma of Molecular Biology
DNA -> RNA -> Protein
Flow of genetic information in cells from DNA to protein
True for eukaryotes and prokaryotes
DNA Replication
DNA replication is the process by which a genome produces an extra copy of itself
- DNA used as a template to make more DNA
- Essential part of cell division
Eukaryotic DNA Replication
Humans, fungi, parasites, plants
Occur in the NUCLEUS of the cell
During S phase (interphase) of the cell cycle
Super-coiled double stranded DNA is unwound & separated by DNA helicase
Helicase
Binds at origin and unwinds and separates DNA strands
DNA replication always occurs in the 5’ to 3’ direction
Leading Strand - continuously replicated
Lagging Strand - Okazaki fragments
Primer
For both fragments, a short piece of RNA called a primer is required as a starting point for the replication process
DNA Polymerase
Walks along the template strand, adding complementary nucleotides to the new strand in the 5’ to 3’ direction
DNA Ligase
Fills in any gaps in the newly replicated strand
Result of Eukaryotic DNA Replication
Two DNA molecules consisting of one new and one old chain of nucleotides (semi-conservative)
-Winds back up into double helix
Telomeres
To prevent the loss of genes as chromosome ends wear down, the tips of eukaryotic DNA have specialized caps called telomeres
- Hundreds of repeats of the same short DNA sequence
- Telomeres also grow shorter in length with each progressive cycle
- Some cells can reverse telomere shortening by expressing telomerase
Bacterial DNA Replication
Conceptually like eukaryotic DNA replication (helicase, primers, DNA polymerase) – HOWEVER, takes place in the CYTOSOL, not nucleus
Both use semi-conservative replication using leading & lagging strand
DIFFERENCES: eukaryotic cell has 25 x MORE DNA than a prokaryotic cell!!
Prokaryotic DNA Replication
- Prokaryotes (bacteria) have only one origin of DNA replication
- DNA polymerase III is the primary enzyme involved in replicating DNA
- Faster rate in prokaryotic cells
- No telomeres (circular genomes)
- Short replication in prokaryotes occurs almost continuously, but eukaryotic cells only undergo DNA replication during the S-phase of the cell cycle
- Bacteria require the enzyme DNA gyrase to resolve supercoils as its circular genome unwinds
Bacterial Replication - Theta Replication
directional from a single origin site
-Following bacterial replication, the resulting circular genomes are concatenated (interlocked) - topoisomerase IV, cuts and reseals the circular chromosomes
Plasmids - Rolling Circle Replication
Plasmids replicate primarily through the rolling circle method
Special enzymes encoded by plasmids (RepA) nick the outer strand of plasmid circular dsDNA
This nick allows DNA polymerase to bind the intact stand which is used as a template
As elongation of this new strand begins, the nicked strand is displaced
The displaced strand is then coped into dsDNA
Method also used by human viruses that have circular dnDNA and bacteriophage
DNA Virus Replication
Viruses may have linear or circular genomes within their capsids
Viral DNA genomes may
-Be used directly as a template with or without association with host nuclear factors
-Linear DNA that circulates in the nucleus is called an “episome” - primarily rolling circle replication
-Integrate into the host genome, propagated with each cell cycle, transformed phenotype
Transcription
Process of using DNA as a template to make RNA
Three types of RNA
rRNA = structural components of ribosomes (nucleolus) mRNA = provides the blueprint for making proteins (transcript) tRNA = uses the blueprint to incorporate the correct amino acids into the growing polypeptide (protein) chain
Transcription requires a __________
DNA-dependent RNA polymerase
Process regulated by transcription factors – proteins that bind to promoter regions upstream of genes initiate or repress transcription
Where does transcription occur?
Eukaryotes = nucleus
Prokaryotes = cytoplasm
Viruses use host ribosomes & often carry proteins that regulate transcription
Introns
Intron = region of mRNA that do not code for proteins
- Are NOT found in prokaryotes
- Simple single celled eukaryotes have few introns compared to more complex eukaryotes
- Some viral mRNAs contain introns that need to be spliced out before translation occurs
Alternative Splicing
In eukaryotes, some genes also contain EXONS which can be differentially spliced out
-The resulting transcripts can code for multiple proteins - increased diversity of genetic output
AS can be found in some viruses (retroviruses like HIV-1 reproduce as one long RNA molecule that is alternatively spliced to generate transcripts (and proteins) of varying types)
Translation
RNA -> Protein Occurs at the ribosome in the cytoplasm Two subunits Eukaryotes: 60S + 40S = 80S Prokaryotes: 50S + 30S = 70S
Ribosome reads the mRNA transcript three nucleotides at a time – CODON
Stop codon is detected, peptide chain released
Translation of Viral Proteins
Viruses utilize the ribosomes of the cells they infect to make their proteins
Viral mRNA may also be translated into large polyproteins that are trimmed down into other functional proteins via the action of viral proteases