7. Bacteriophages 1 Flashcards

(73 cards)

1
Q

What is the unique thing about MS2 phage?

A

protein translation

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2
Q

what is the unique thing about phiX174 phage?

A

rolling circle replication of DNA

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3
Q

what is the unique thing about T7 phage?

A

transcription coordinated with entry

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4
Q

what is the unique thing about lambda phage?

A

prophage (integrates phage DNA into bacterial host DNA) and complex regulation of gene expression

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5
Q

which domains of life are infected by viruses?

A

all 3! bacteria, archaea, and eukaryotes

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6
Q

what are the 3 similarities btwn bacteria and archaea?

A
  1. unicellular
  2. circular DNA
  3. no nucleus
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7
Q

what are 9 unique characteristics that archaea have but bacteria don’t?

A
  1. cell walls made of impermeable S layer proteins
  2. special phospholipids
  3. translation initiation mimics eukaryotes
  4. uses methionine (not N-formyl methionine) to start translation
  5. histone-like proteins to package chromosomes
  6. complex RNA pol machinery
  7. DNA replication machinery mimics eukaryotes
  8. distinct rRNA sequences
  9. live in extreme environments
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8
Q

what is the morphology of viruses of archaea?

A

UNUSUAL!
- lemon
- droplet
- bottle
etc

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9
Q

what type of genomes do viruses of archaea have?

A

dsDNA except 1 has ssDNA

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10
Q

describe the envelopes of viruses of archaea?

A

MOST have internal or external lipid envelopes

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11
Q

what does it mean for viruses of archaea to be temperate?

A

temperate = LATENT –> they integrate their genome into host cell DNA without killing the cell

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12
Q

describe the DNA polymerase of viruses of archaea?

A

many don’t have identifiable DNA polymerase gene

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13
Q

what virus is this?

A

sulfolobus spindle-shaped virus 1 (SSV1)

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14
Q

what virus is this?

A

acidianus two tailed virus (ATV)

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15
Q

what virus is this?

A

acidianus bottle-shaped virus (ABV)

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16
Q

what virus is this?

A

sulfolobus neozealandicus droplet-shaped virus (SNDV)

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17
Q

why do we not need to worry about viruses of archaea?

A

we don’t go to the extreme environments where archaea live

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18
Q

when were phages discovered? by who?

A

Twort and d’Herelle discovered them in 1915/1917

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19
Q

when were phages first visualized?

A

in 1940

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20
Q

what was found in 1969? by who?

A

Delbruck, Hershey, and Luria found how phages replicate DNA and express genes

i.e. using phages, found that DNA was genetic material bc DNA alone could dictate production of new phages

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21
Q

what was found in 1978? by who?

A

Arber, Nathans, and Smith discovered restriction enzymes and how they could be used to open a vector and insert DNA for cloning

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22
Q

how do bacteria use restriction enzymes? explain

A

bacteria use them as a defense mechanism against phages

restriction enzymes target a small DNA sequence –> this sequence is modified in bacteria but remains the same in phages so the enzymes can cleave phage DNA

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23
Q

when were viruses sequenced? what type of tools were used + 3 examples?

A

MS2 - 1976
phiX174 - 1977
lambda - 1982 using PHAGE-DERIVED TOOLS –> restriction enzymes, T4 DNA ligase, M13 vectors

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24
Q

how do we use phage-derived tools?

A

to sequence everything

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25
what was found in the 1990s?
there is an abundance of phages in our ecosystem that have co-evolved with bacteria via gene transfer
26
besides restriction enzymes, what is another defense mechanism of bacteria against phages? how does it work?
CRISPR acts as bacterial immune system memory --> phage DNA is inserted into host DNA so the bacteria can remember which phages were there to attack upon next infection
27
how do we use CRISPR in labs?
to edit genes
28
what is phage therapy used for?
to target bacterial infection
29
where does the word bacteriophage come from?
bacteria + phagein (to devour)
30
what type of molecule are bacteriophages?
obligate intracellular parasites
31
which type of bacteria do bacteriophages infect?
all natural populations of bacteria
32
what are the most widely distributed and diverse entities in the biosphere? how many
bacteriophages --> 10^31
33
do all bacteriophages have the same structure?
no
34
what is the structure of T4 phages?
head, contractile tail, long fibers
35
what is the structure of lambda phages?
head, long non-contractile flexible tail, short fibers
36
what is the structure of T7 phages?
no tail, short fibers, head
37
what is the structure of phiX174 phages?
only head
38
What is the structure of M13 phage?
no head only fiber (ebola similar shape)
39
what are the 3 possible type of genomes in bacteriophages?
1. ssRNA 2. ssDNA 3. dsDNA
40
what are the 2 families of ssRNA bacteriophages? and examples of each
1. LEVIviridae --> MS2 2. INOviridae --> f1, fd, M13
41
what is the family of ssDNA bacteriophages? and an example
MICROviridae --> phiX174
42
what are the 2 families of dsDNA bacteriophages? and examples of each
1. PODOviridae --> T7 2. SIPHOviridae --> lambda
43
what is unique about dsDNA bacteriophages?
they have tails
44
do bacteriophages have envelopes?
no
45
describe MS2 genome
(+) ssRNA, 4kb
46
how many maturation proteins are in the MS2 phage?
1
47
how many coat proteins are in the MS2 phage?
180 copies
48
describe MS2 entry and lifecycle in bacterial cell (5 steps)
1. virus binds to sex pilus 2. viral RNA enters the cell 3. ssRNA translates into phage proteins 4. ssRNA: (+) --> (-) to make more (+) 5. assembly
49
how many viral phage genes are in MS2? what are they?
1. maturation 2. coat 3. lysis 4. replicase
50
what proteins are in the MS2 viral particle? which are not?
maturation and coat protein in MS2 particle lysis and replicase are not
51
what are the 5 general steps of MS2 protein synthesis?
1. coat protein produced upon entry 2. allows replicase and lysis proteins to be produced 3. replicase binds to start codon of coat gene to block coat protein synthesis, copies (+) RNA --> (-) --> (+) to make more genome 4. the new (+) RNA folds to allow synthesis of maturation protein 5. MORE coat proteins produced and form dimers that stop replicase production and initiate phage assembly
52
what is the role of replicase protein?
amplify phage genome
53
what is the role of MS2 coat protein on 1st production and 2nd production?
1st: allows production of replicase and lysis proteins 2nd: shut down replicase protein and initiate assembly of phage particles
54
how does the production of MST coat protein allow for production of replicase and lysis proteins?
coat protein start codon is easily accessible but lysis and replicase start codons are hidden: 1. host ribosomes can recognize start codon of coat protein to translate it 2. once coat is translated, lysis and replicase hairpins open and expose their start codons
55
how does folding of RNA regulate the translation of maturation protein?
when the RNA is folding up, it has an intermediate structure for a short time where the ribosome can access the start codon and make a few maturation protein then the mature RNA is folded up, hiding the ribosome entry site for the maturation protein
56
what does replicase do? what helps replicase do its job?
replicase binds to the start of the coat gene to shut down coat protein translation replicase is associated with 3 HOST proteins
57
what are the 3 host proteins associated with replicase? and what do they each do?
1. S1 protein in the small ribosomal subunit --> directs replicase to the start of coat gene 2. EF-Tu translational elongation factor --> with GTP, helps replicase initiate RNA synthesis 3. EF-Ts translational elongation factor --> recycles EF-Tu/GDP to EF-Tu/GTP
58
how do we use MS2/RNA binding to monitor RNA?
add MS2 to 3' end of gene and fuse with GFP MS2 binds to RNA and when RNA is exported to the cytoplasm, GFP indicates where RNA is located
59
what type of virus is phiX174?
microvirus
60
what type of genome does phiX174 have?
ssDNA
61
does phiX174 have its own polymerase?
no --> uses host polymerase
62
describe phiX174 entry into the cell (2 steps)
1. capsid interacts with sugar residues (ex. glucose) in LPS of bacteria 2. H protein in spike penetrates cell wall then forms channel for DNA to be delivered thru viral spikes
63
what proteins are in phiX174 spikes?
G and H proteins
64
what allows H protein to mediate the penetration step?
H protein has an N-terminal transmembrane helix
65
what is the first stage of phiX174 genome replication?
circular (+) ssDNA uses host proteins to start DNA synthesis with primosome --> host DNA pol makes circular dsDNA
66
what is the second stage of phiX174 genome replication?
viral A protein cleaves circular dsDNA to make exposed 3' end that acts as a primer to make ssDNA --> elongates --> new ssDNA is cleaved out and circularized to make new (+) ssDNA
67
how many promoters does the phiX174 genome use?
3
68
how many terminators does the phiX174 genome use?
4
69
what 4 factors control viral gene expression of phiX174?
1. transcription strength of promoters 2. stability of viral RNA 3. termination efficiency of terminators 4. ribosome binding site
70
if there are only 3 promoters, how can phiX174 express more than 3 genes?
promoter stops at different sites to control the amount of mRNA and therefore the amount of protein
71
what are the 2 scaffolding proteins expressed by phiX174? what is their role for phiX174 phage?
protein B and protein D they help with assembly, then removed by protease to allow complete maturation and become infectious
72
why is a provirion not infectious?
even though it has all its proteins, a provirion still has scaffolding proteins
73
what does the assembly of phiX174 start with?
starts with assembled units, not monomer