Molecular Genetics Flashcards

1
Q

Nucleotide

A
  • ribose sugar, nitrogenous base, and
    phosphate group.
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2
Q

Nucleoside

A
  • ribose sugar and nitrogenous base.
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3
Q

DNA is a polymer of ______ that have
hydrogen on the ribose sugar’s 2’ carbon. RNA is a
polymer of nucleotides that have ______ groups
on the ribose sugar’s 2’ carbon. This is the reason
DNA is called deoxyribonucleic acid, while RNA is
called ribonucleic acid.

A

1) nucleotides
2) OH-

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4
Q

Since ______ have more hydrogen bonds, a
______ temperature is needed to break DNA
strands.

A

1) G-C bonds
2) higher

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5
Q

Nucleosomes

A
  • are complexes of DNA wrapped
    around histone proteins. Each nucleosome has
    nine histones total. The central core contains two
    of each histone H2A, H2B, H3 and H4. On the
    outside, a single histone, H1, holds the DNA in
    place.
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6
Q

Chromatin

A
  • refers to the overall packaging of DNA
    and histones.
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7
Q

2 Types of Chromatin include

A

1) Euchromatin
2) Heterchromatin

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8
Q

Euchromatin

A
  • nucleosomes are “loosely
    packed”, so DNA is readily accessible for
    transcription.
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9
Q

Heterochromatin

A
  • nucleosomes are “tightly
    packed”, so DNA is mostly inactive.
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10
Q

Histones are _____ charged while DNA is
_______ charged, allowing proper binding.

A

1) positively
2) negatively

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11
Q

Acetylation

A

-of histones removes positive charges,
relaxing DNA-histone attractions and allowing for
more transcription to happen.

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12
Q

Deacetylation

A
  • of histones increases positive
    charges, tightening DNA-histone attractions and
    decreasing transcription.
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13
Q

Methylation

A
  • of histones adds methyl groups, either increasing or decreasing transcription.
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14
Q

An _______ is required to initiate DNA replication where the DNA strands first separate. Organisms with ______ DNA such as bacteria have a single origin of replication while organisms with ______ DNA such as humans have multiple origins of replication.

A

1) origin of replication
2) circular
3) linear

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15
Q

DNA undergoes _______,
where each new double helix produced by
replication has one “new” strand and one “old”
strand.

A

1) semiconservative replication

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16
Q

DNA is ______, meaning that the _______
(terminal phosphate group) of one strand is
always next to the _____ (terminal hydroxyl
group) of the other strand and vice versa.

A

1) antiparallel
2) 5’ end
3) 3’ end

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17
Q

Steps of Replication

A

1) Initiation
2) Elongation
3) Termination

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18
Q

Initiation

A
  • creating origins of replication at
    A-T rich segments of DNA because A-T bonds
    only have two hydrogen bonds and are easier
    to split apart.
  • a promoter sequence (aka
    promoter) next to the gene attracts RNA
    polymerase to transcribe the gene.
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19
Q

Elongation

A
  • producing new DNA strands using different types of enzymes.
  • transcription bubble forms and RNA polymerase travels in the 3’ → 5’ direction
    on the template strand. However, it extends
    RNA in the 5’ → 3’ direction.

Involves:
Helicase
Single-strand binding proteins
topoisomerase
Primase
sliding clamp proteins
DNA polymerase(s)
leading strand
lagging strand
DNA ligase

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20
Q

Helicase

A

-unzips DNA by breaking hydrogen bonds between strands, creating a replication fork. As it unzips the strands, helicase leads to supercoiling (tension ahead of the replication fork).

-Separates complementary strands at the replication fork.

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21
Q

Single-strand binding proteins

A
  • bind to uncoiled DNA strands, preventing
    reattachment of the strands to each other.
  • Proteins that prevent
    the two strands from
    coming back together
    after separate.
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22
Q

Topoisomerase

A
  • nicks the DNA double
    helix ahead of helicase to relieve built-up
    tension and supercoiling.
  • Relaxes the DNA double
    helix from the tension
    and supercoiling the
    opening helix is creating.
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23
Q

Primase

A
  • Primase places RNA primers at the origin
    of replication to create 3’ ends for nucleotide addition.
  • Provides a 3’ hydroxyl
    group for DNA
    polymerase to attach
    new nucleotides to.
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24
Q

Sliding clamp proteins

A
  • hold DNA polymerase onto the template strand.
  • Helps to hold DNA
    polymerase to the
    template strand.
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25
DNA polymerase
- adds free nucleoside triphosphates to 3’ ends. DNA polymerase can only add nucleotides onto an preexisting 3’ hydroxyl group provided by primase. - The class of enzymes that extends DNA in the 5’ to the 3’ direction. Several have proofreading capabilities that allow them to catch synthesis errors.
26
The leading strand
-is produced continuously because it has a 3’ end that faces the replication fork.
27
The lagging strand
- is produced discontinuously because its 3’ end is facing away from the replication fork. Thus, many RNA primers are needed to produce short DNA fragments called Okazaki fragments.
28
A different _______ replaces RNA primers with DNA.
1) DNA polymerase
29
DNA ligase
-glues separated fragments of DNA together. - GLues together separate pieces of DNA.
30
Termination
- replication fork cannot continue, ending DNA replication. - a termination sequence (aka terminator) signals to RNA polymerase to stop transcribing the gene.
31
Telomeres
- are noncoding, repeated nucleotide sequences at the ends of linear chromosomes. They are necessary in eukaryotes because when the replication fork reaches the end of a chromosome, a small segment of DNA from the telomere is not replicated and lost (no RNA primer is present to help produce another Okazaki fragment).
32
To review, the _____ checkpoint regulates cell cycle transition from the _____ into the S phase, checking for favorable conditions to grow. If _______, the cell will remain in _______ and will not enter the S phase for DNA replication.
1) G1/S 2) G1 phase 3) unfavorable 4) G0
33
Telomerase
- Adds repetitive DNA to the ends of eukaryotic chromosomes, which prevents critical information from being lost. -Telomerase is an enzyme that extends telomeres to prevent DNA loss.
34
Genes
- are instructions within DNA that code for proteins. However, they must first be transcribed into RNA before being translated into proteins. In a gene, the promoter region comes first, then the gene operator, then the gene.
35
Specifically, DNA undergoes transcription to produce single-stranded __________.
1) mRNA
36
Because ______ do not have membrane-enclosed nuclei, both transcription and translation occur simultaneously in the cytosol. RNA polymerase opens up DNA, forming a transcription bubble.
1) Prokaryotes
37
Before transcription can occur, a _______ combines with _______ to form RNA polymerase holoenzyme, giving it the ability to target specific DNA promoter regions.
1) sigma factor 2) prokaryotic core RNA polymerase
38
There are two types of termination in bacteria:
1) Rho independent termination 2) Rho-dependent termination
39
Rho independent termination
— a termination sequence is reached and the RNA transcript folds into a hairpin loop → RNA polymerase falls off and transcription ends.
40
Rho-dependent termination
— A Rho protein binds to the RNA transcript, moving 5’ → 3’ to catch up and displace RNA polymerase, ending transcription.
41
operon
is a group of genes that function as a single unit that is controlled by one promoter. The operator region is present near the operon’s promoter.
42
To regulate the promoter, _______ bind to the operator regions, while _______ bind to the promoter sites. (prokaryotes)
1) repressor 2) activator
43
The lac operon
is an inducible operon (it must beinduced to become active). LacZ, lacY, and lacA are the three genes contained within the lac operon that encode proteins required for lactose metabolism. The lac operon will only be induced when glucose is not available as an energy source, so lactose must be used.
44
The lac repressor protein
is the first way that the lac operon is controlled. This protein is constitutively expressed (always on). Thus, the lac repressor protein is always bound to the operator, blocking transcription. However, when lactose is present it is converted to allolactose. Allolactose binds directly to the repressor and removes it from the operator, allowing transcription to occur.
45
cAMP levels and catabolite activator protein (CAP)
- are the second level of lac operon regulation. cAMP levels are inversely related to glucose levels, so when glucose is low, cAMP is high. cAMP binds to catabolite activator protein (CAP), which then attaches near the lac operon promoter to help attract RNA polymerase, promoting transcription. Glucose and lactose present: - Moderate transcription. Repressor not bound and CAP not bound. Glucose and lactose absent: - No transcription. Repressor is bound and CAP is bound. Glucose present and lactose absent: - No transcription. Repressor is bound and CAP is not bound. Glucose absent and lactose present: - High transcription. Repressor not bound and CAP is bound.
46
trp operon
- is responsible for producing the amino acid tryptophan. It is known as a repressible operon because it codes for tryptophan synthetase and is always active unless the presence of tryptophan in the environment represses the operon.
47
______ binds to the trp repressor protein, which then attaches to the ______ on the trp operon to prevent tryptophan production. Thus, this is the first level of trp operon regulation. When tryptophan is not present in the environment, the trp operon will undergo transcription because the trp repressor protein will be _______. (prokaryotes)
1) Tryptophan 2) operator 3) inactive
48
Unlike in prokaryotes, eukaryotic transcription occurs in the ______ and uses ________ to transcribe most genes.
1) nucleus 2) RNA polymerase II
49
Transcription factors
- are needed in eukaryotes to help RNA polymerase bind to promoters. The TATA box is a sequence in many promoters that transcription factors can recognize and bind to.
50
Enhancers
- are DNA sites that activator proteins can bind to; they help increase transcription of a gene.
51
Silencers
- are DNA sites that repressor proteins can bind to; they decrease transcription of a gene.
52
Enhancers and silencers can be far ______ or _______ from the gene, so DNA from these sites are thought to loop around to colocalize with RNA polymerase.
1) upstream 2) downstream
53
The poly A signal
- is located within the terminator sequence and stimulates polyadenylation (addition of adenine nucleotides to the 3’ end of the mRNA).
54
Exonucleases
- are enzymes that cleave nucleotides from the polynucleotide chain at the ends of the chain. Exonuclease activity only results in sticky ends.
55
Endonucleases
- are enzymes that cleave nucleotides from the polynucleotide chain from the inside of the polynucleotide chain. Endonuclease activity can result in either sticky or blunt ends.
56
Post-transcriptional modification (PTM) (Eukaryotes)
- describes the conversion of pre-mRNA into processed mRNA, which leaves the nucleus.
57
There are 3 main types of PTM
1) 5' capping 2) Polyadenylation of 3' end 3) Splicing out introns
58
5’ capping
- 7-methylguanosine cap is added to the 5’ end of the mRNA during elongation, protecting the mRNA from degradation.
59
Polyadenylation of the 3’ end
- addition of the poly A tail to the 3’ end to prevent degradation by exonucleases.
60
Splicing out introns
- introns are stretches of noncoding DNA that lie between regions of coding DNA (exons). Splicing refers to removing introns from pre-mRNA using spliceosomes. “Splice signals” present within introns signal to the spliceosome where to cut.
61
Alternative splicing
- describes a single pre-mRNA having multiple possible spliced mRNA products. Thus, the same pre-mRNA can produce many different proteins.
62
miRNAs (micro RNA)
-are small RNA molecules that silence mRNA expression as a method of post-transcriptional gene regulation by base-pairing with parts of sequences on the mRNA transcript that inhibits their translation.
63
snRNAs (small nuclear RNA) and proteins
- make up the functional part of a spliceosome and are collectively referred to snRNPs (small nuclear RiboNucleic Proteins).
64
_____ and _____ are important players in translation, the process of converting mRNA into protein products.
1) ribosomes 2) tRNA
65
Difference in ribosome makeup: Eukaryotes vs Prokaryotes
1) Eukaryotes - small (40S) and large (60S) subunits form a 80S ribosome. They are composed of rRNA (ribosomal RNA) and proteins. The subunits are made in the nucleolus and assembled once they are exported to the cytosol. 2) Prokaryotes - small (30S) and large (50S) subunits form a 70S ribosome. They are also composed of rRNA and proteins, but are assembled together in the nucleoid.
66
A codon
- is a group of three mRNA bases (A, U, G, or C) that code for an amino acid or terminate translation. There are 64 codon combinations total but only 20 amino acids, so degeneracy is present (multiple codons code for the same amino acid).
67
Start Codon
- AUG (Methione)
68
Stop codons
- UAA, UAG, UGA (end translation, do not code for any amino acid)
69
An anticodon
- is a group of three tRNA bases (A, U, G, or C) that base pairs with a codon. Each tRNA carries an amino acid to be added to the growing protein.
70
Aminoacyl-tRNA
- refers to a tRNA bound to an amino acid.
71
Aminoacyl-tRNA synthetase
- is the enzyme that attaches an amino acid to a specific tRNA using the energy from ATP.
72
Ribosomal Binding sites for tRNA
1. A site - A for aminoacyl-tRNA, which first enters at this site. 2. P site - P for peptidyl-tRNA, which carries the growing polypeptide. 3. E site - E for exit site. The tRNA from the P site is sent here and released from the ribosome.
73
The ribosome catalyzes the formation of a _____ between the polypeptide in the P site and the newly added amino acid in the A site. Afterwards, the polypeptide is transferred to the _______ and the ribosome shifts one codon down the mRNA. The _____ will now be empty and ready to accept another aminoacyl-tRNA. The tRNA from the ______ will be transferred to the _____ and will leave the ribosome. During ______ (occurs in which the tRNA molecule at the A site moves to the P sit), and the tRNA at the P site moves to the E site (A → P → E)
1) peptide bond 2) A site’s tRNA 3) A site 4) P site 5) E site 6) translocation
74
Chaperonins
- Specialized proteins found in both eukaryotic and prokaryotic organisms and function in assisting newly synthesized polypeptides to fold into their correct shape.
75
A DNA mutation
- is a heritable change in the DNA nucleotide sequence that can be passed down to daughter cells.
76
2 main types of DNA mutations:
1) Base substitution (point mutation) 2) Frameshift mutation
77
Base substitutions (point mutations)
- one nucleotide is replaced by another. - includes: silent mutation, missense mutation, and nonsense mutation
78
Silent mutations
- no change in amino acid sequence. Due to “third base wobble”, mutations in the DNA sequence that affect the third base of a codon can still result in the same amino acid being added to the protein. Relies on the degeneracy (redundancy) of translation.
79
Missense mutations
- single change in amino acid sequence. Can either be conservative (mutated amino acid similar to unmutated) or non-conservative (mutated amino acid different from unmutated).
80
Nonsense mutations
- single change in amino acid sequence that results in a stop codon. Results in early termination of protein
81
Frameshift mutations
- are mutations that result in a shift in the reading frame, changing the way the mRNA transcript is read. - there are 2 types: insertions and deletions
82
Insertions
- adding nucleotides into the DNA sequence - can shift the reading frame.
83
Deletions
- removing nucleotides from the DNA sequence - can shift the reading frame.
84
A null mutation
- can also occur, a null (non-functional) allele is produced that lacks the function of the normal, wild-type allele.
85
Factors that contribute to DNA mutations:
● DNA polymerase errors during DNA replication. ● Loss of DNA during meiosis crossing over. ● Chemical damage from drugs. ● Radiation ● Transposons (jumping genes) - DNA sequences in prokaryotes and eukaryotes that can move and integrate into different places in the genome and cause mutations.
86
Factors that prevent DNA mutations:
● DNA polymerase proofreading by DNA polymerase. ● Mismatch repair machinery that checks uncaught errors. ● Nucleotide excision repair that cuts out damaged DNA and replaces it with correct DNA using complementary base pairing.
87
Chromosomal mutations
- occur and affect the entire chromosome rather than individual nucleotides. - There are four types: 1) duplication 2) translocation 3) deletion 4) inversion
88
Duplication
- A region of DNA is duplicated, resulting in a larger chromosomal arm and an atypical banding pattern.
89
Translocation
- A piece of one chromosome breaks off and attaches to another chromosome. Translocation increases chromosomal arm length and results in an abnormal banding pattern. This is the only mutation that affects both chromosomes.
90
Deletion
- A portion of the chromosome is deleted, resulting in a shorter chromosomal arm.
91
Inversion
- A portion of the chromosome becomes inverted on the arm of the chromosome. Results in an abnormal banding pattern, but does not affect the length of the chromosome.
92
______ are not living because they must infect living cells to multiply.
1) Viruses
93
The ______ is a viral protein coat that is made of subunits called _______. Some viruses also have a phospholipid envelope that they pick up from the host cell membrane.
1) capsids 2) capsomere
94
In order for an infection to continue to spread, viruses undergo ______ to create new viruses that can further infect other cells/hosts.
1) Viral replication
95
The viral replication cycle is as follows:
1. Attachment - binding of a virus to host cell 2. Penetration - virus crosses through the host’s cell membrane. 3. Uncoating - viral capsid is removed and degraded by host enzymes. 4. Synthesis - components of viral capsid are manufactured. 5. Assembly - viral capsid components assemble to form the viral capsid. 6. Release - last step of viral replication, fully assembled viruses are released.
96
Two viral life cycle types:
1) Lysogenic Cycle 2) Lytic Cycle
97
Lysogenic cycle
- virus is considered dormant because it inserts its own genome into the host’s genome and does not harm the host. Each time the host genome undergoes replication, so does the viral genome.
98
Lytic cycle
- virus takes over host to replicate and does cause harm to the host. The viral particles produced can lyse the host cell to find other hosts to infect.
99
Viruses can ____ between the lysogenic and lytic cycles. For example, _____ can stimulate a virus in the lysogenic cycle to replicate and enter the lytic cycle.
1) switch 2) favorable conditions
100
Retroviruses (eg. HIV)
- have an RNA genome that infects host cells. They contain an enzyme called reverse transcriptase, which converts their RNA into cDNA (complementary DNA). The cDNA can integrate into the host genome and enter the lysogenic cycle.
101
_____ are asexual and divide by ______, so they only receive genes from one parent cell and do not increase genetic diversity through reproduction. ______ are bacteriophage genomes that have been integrated into the host genome.
1) asexual 2) binary fission 3) prophages
102
Bacteria must increase genetic diversity through ______, which describes the transfer of genes between individual organisms.
1) horizontal gene transfer
103
There three methods of horizontal gene transfer:
1) conjugation 2) transformation 3) transduction
104
Conjugation
- bacteria use a cytoplasmic bridge called a pili to copy and transfer a special plasmid known as the F plasmid (fertility factor). If a bacteria contains an F plasmid, it is referred to as F+. If not, it is referred to as F-. To review, plasmids are circular DNA pieces that are independent from a bacteria’s single circular chromosome.
105
Transformation
- - bacteria take up extracellular DNA. Bacteria are referred to as competent if they can perform transformation. Electroporation is the process of using electrical impulses to force bacteria to become competent.
106
Transduction
- viruses transfer bacterial DNA between different bacterial hosts. This occurs when a bacteriophage enters the lysogenic cycle in its host and carries bacterial DNA along with its own genome upon re-entering the lytic cycle.
107
Quorum sensing
- mechanism of communication by cells, regulating the release of signaling molecules that affect microbial metabolism and gene expression. Dependent on cell density.
108
Bacteria can also contain ______, which are extrachromosomal pieces of DNA in the form of a plasmid that contains antibiotic resistance genes.
1) R-factors