Chapter 9 Flashcards

(40 cards)

1
Q

Genomic analysis shows that microbes undergo extensive gene loss and gain

result of?

A

20% of E. coli genome may have originated in other microbes

horizontal gene transfer, recombinations, and a variety of mutagenic and DNA repair strategies

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2
Q

Horizontal gene transfer

A

movement of genes between species or genera

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3
Q

vertical gene transfer

A

generational passing of genes from parent to offspring (cell division)

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4
Q

transformation

A

importing DNA into cells
• At least 82 known naturally competent (capable of importing DNA) species
• Gram-positive (Streptococcus, Bacillus)
• Gram-negative (Haemophilus, Nisseria)
Requires specific protein complexes called transformasomes

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5
Q

• Why is natural transformation useful?

A
  • Use as ”food”
  • DNA repair
  • Influence evolution – species adjust to new envir. by new genes.
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6
Q

Gram-positive – growth phase dependent competence

A
  • Triggered by quorum sensing
  • Each cell secretes a small peptide called competence factor (CF) – unique to each species
  • CF increases as population increases
  • Above a certain concentration CF activates a phosphorylation cascade that activates transcription of transformasome
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7
Q

Gram-negative

A

•no CF-always competent (Nisseria) or become comp. when starved (Haemophilus)
•Have a similar transformasome
•Outer membrane barrier
-Neisseria use pilus assembly-when pilus disassembles it drags DNA into cell
-Pilus naturally assembles and disassembles during growth

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8
Q

Transformation is species and sequence specific

A

limits exchange between genera

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9
Q

conjugation

A

cell-cell contact w sex pilus sticking out from a donor cell

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10
Q

Hfr strain

A

high-frequency recombination strain – F-

factor integrated into chromosome

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11
Q

Generalized transduction

A

take gene from donor cell to recipient
• Uses rolling circle-makes long copy of dna (concatemers)
• pac site to cut them into indiv. pieces

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12
Q

Specialized transduction

A

transfer only a few closely linked genes b/w cells
• Example (E. coli)
• integrates into host chromosome(lysogenic)
• Improper excision by host recombination enzymes – take host genes adjacent to attachment site

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13
Q

What is the fate of new DNA that has entered the cell?

A
  • Plasmid capable of autonomous replication
  • May be incorporated into chromosome
  • Degraded by nucleases –seen as foreign DNA bc lack of methylation or by CRISPR
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14
Q

Generalized recombination

A

two recombining molecules w significant homology (crossing over in sections of dna)

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15
Q

Site specific recombination

A

little sequence homology – requires short (10-20 bp) sequence recognized by recombination enzyme

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16
Q

Why is recombination advantageous?

A
  • DNA repair mechanism
  • Cells w/ damaged chromosomes use DNA donated by others from same species to repair damaged genes
  • “Self-improvement” program – samples genes from other organisms to enhance fitness of cell
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17
Q

Mutation

A

• change in base sequence and failure for cell to repair the change

18
Q

point mutation

A

change in a single nucleotide

19
Q

silent mutation

A

does not change AA sequence. lys–>lys (degenerate codon)

20
Q

Missense mutation

A

changes aa sequence

21
Q

Loss-of-function mutation

A

decreases or eliminates protein activity

22
Q

gain-of-function mutation

A

increases or gain new activity

23
Q

knockout mutation

A

eliminates function

24
Q

Frameshift mutation

A

insertion or deletion of 1-2 bp (not multiple of 3). ribosome reads wrong triplets. sometimes stop codon.

25
mutagens | ex. uv light
chemical agents that can damage DNA | for pyrimidine-creates pyrimidine dimer that blocks replication/transcription
26
spontaneous mutations
• Rare bc DNA repair/proofreading is efficient • Tautomeric shifts- C binds to A • Deamination reactions – C change to U • Damage by reactive oxygen species (H2O2, superoxide/hydroxyl radicals) -interfere w/ polymerases and stops replication/transcription
27
Methyl mismatch repair
deoxyadenosine methylase (Dam) –methylates GATC • Only parent strand methylated • Mut proteins-cut out incorrect base, DNA Pol I fills in correct base
28
DNA repair – Error proof repair | • Photoreactivation
Photolyase binds pyrimidine dimer and cleaves cyclobutane ring linking damaged nucleotides
29
DNA repair – Error proof repair | • Nucleotide excision repair
* removes 12-13 nucleotides | * DNA Pol I repairs gap
30
DNA repair – Error proof repair | • Base excision repair
``` • Glycosylase clip damaged bases -U fromC deamination -Deamination of A to hypoxanthine • Cuts out single base • Endonuclease cleaves the backbone • DNA Pol I repairs strand ```
31
• E. coli’s genome is rife with
genomic islands, inversions, deletions, paralogs and orthologs
32
Transposable elements
enzyme that copies sequence from one dna into anther (tranasposase enzyme makes this happen) • Not autonomous – cant exist outside of larger DNA molecule
33
insertion sequences (FOR TRANSPOSABLE ELEMENTS)
simple transposable element (700- 1500 bp)
34
Transposons
carry other genes plus those required for transposition
35
transposition
moving a transposable element b/w DNA
36
stop codons
uaa, uga, uag
37
F' factor
takes some chromosomal dna
38
DNA repair – Error proof repair | Recombinational repair
•both strand/one strand is damaged -DNA Pol III skips over damaged regions • RecA binds gap and w/ piece of undamaged DNA • Damage can be repaired by other mechanisms
39
DNA repair – Error proof repair | • SOS (“save our ship”) repair
extensive damage • RecA binds ssDNA (stimulates autodig. of LexA repressor) “sloppy ”polymerases w/ NO proofreading- insert whatever is available
40
DNA repair – Error proof repair | Nonhomologous end joining
Double stranded breaks are | dangerous. Ku and LigD recombine them. can be errors.