3- Viral pathogens: classification, biology, diseases - I Flashcards

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1
Q

Define what a virus is

A

➝An infective agent that typically consists of a nucleic acid molecule in a protein coat, is too small to be seen by light microscopy, and is able to multiply only within the living cells of a host- obligate cellular parasites.

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2
Q

What are the 4 types of viral genome?

A

➝ ssRNA
➝ dsRNA
➝ ssDNA
➝ dsDNA

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3
Q

What are the 2 types of DNA genome?

A

➝DNA genomes can be linear or circular.

➝Double-stranded genomes have complementary base pairing

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4
Q

What are the 2 types of RNA genome?

A

➝RNA genomes can be linear and segmented i.e. more than one RNA per capsid

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5
Q

What are the two ways that viral genomes can encode information?

A

➝ positive or negative

➝ 5’-3’ or 3’5

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6
Q

What is the central dogma of molecular biology?

A

Replication, Transcription, and Translation

➝ DNA ➝ RNA ➝ Protein

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7
Q

Why does the central dogma fall apart?

A

➝ Viruses can go from RNA ➝ DNA

➝using reverse transcriptase

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8
Q

How do viruses use the central dogma to their own advantage?

A

➝DNA viruses- replicate DNA via DNA polymerase
➝Produce RNA from DNA via RNA polymerase
➝RNA viruses replicate RNA via RNA dependent RNA polymerase
➝Retroviruses- produce DNA from RNA via reverse transcriptase

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9
Q

What are the 7 groups that make up the baltimore classification?

A

➝ Group 1 - ds DNA (+/-)
➝ Group 2 - ssDNA (+)
➝ Group 3 - dsRNA (+/-)
➝ Group 4 - ssRNA (+) positive sense
➝ Group 5 - ssRNA (-)
➝ Group 6 ssRNA (+) reverse transcriptase
➝ Group 7 DNA (+/-) reverse transcriptase

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10
Q

What is the structure of HIV?

A

➝ A nucleic acid (two RNA strands)
➝ a protein capsid
➝ a lipid bilayer membrane
➝ proteins on the top that mediate entry into the cell

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11
Q

What does the outer layer of HIV consist of?

A

➝ lipid bilayer with protruding Env spikes

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12
Q

What are Env spikes made from?

A

➝ heterotrimers of SU3TM3

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13
Q

What is inside the envelope of HIV?

A

➝ Gag proteins

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14
Q

How is the conical capsid formed?

A

➝ Matrix associates with the membrane capsid to form the conical capsid

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15
Q

What coats the viral RNA genome in HIV?

A

➝ Nucleocapsid

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16
Q

What does the HIV core contain?

A

➝ two genomic RNA strands (plus strand)
➝ tRNA lys53
➝ ~50 copies of each viral enzyme: PR, RT and IN

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17
Q

What are the three polyproteins that retroviruses synthesize?

A

➝ Gag
➝ Pol
➝ Env

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18
Q

What does Gag code for?

A
➝ group specific antigen
➝ viral core proteins
➝ matrix
➝ capsid
➝ nucleocapsid
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19
Q

What does Pol code for?

A

➝ viral enzymes
➝ protease
➝ reverse transcriptase
➝ integrase

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20
Q

What does Env code for?

A

➝ Envelope glycoprotein : Gp120 surface

➝gp41 transmembrane

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21
Q

What are the 6 accessory/ regulatory proteins to HIV?

A
➝ Tat
➝ Rev
➝ Vif
➝ Nef
➝ Vpu
➝ Vpr
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22
Q

What is the function of Tat?

A

➝ potent activator of viral transcription

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23
Q

What is the function of Rev?

A

➝ mediates unspliced RNA nuclear export

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24
Q

What is the function of Vif?

A

➝ Critical regulator of virus infectivity

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25
Q

What is the function of Nef?

A

➝ Immune modulator, T cell activation, virus spread

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26
Q

What is the function of Vpu?

A

➝ immune modulator, virus release

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27
Q

What is the function of Vpr?

A

➝ Cell cycle, virus nuclear import

28
Q

What proteins does the HIV envelope consist of?

A

➝ Trimer of gp41 and gp120 peptide subunit covered with glycans

29
Q

What proteins does HIV-1 require for entry?

A

➝ CD4 and a chemokine receptor (CCR5/CXCR4)

30
Q

What is the cellular receptor for HIV in humans?

A

➝ CD4

31
Q

How does HIV enter the membrane?

A

➝ The envelope glycoprotein samples the membrane
➝ it sticks to CD4
➝ HIV specific CD4 receptors and co-receptors CCR5 and CXCR4 which also recognise CD4
➝ 6 helix bundle structure is formed
➝ the membrane fusion allows the virus to enter the cell

32
Q

What is the 6 helix bundle ?

A

➝ The structure of the envelope meshing the viral membrane and the cellular plasma membrane

33
Q

What is HIV tropic to?

A

➝ CD4 expressing cells such as T helper cells and macrophages

34
Q

What happens if the body loses helper T cells and macrophages?

A

➝ AIDS

➝ immunodeficiency

35
Q

What does the HIV do while it is travelling to the nuclear membrane?

A

➝ Produces viral DNA via reverse transcription

36
Q

What gives the HIV directionality inside the cell and what is this called?

A

➝ moving down the microtubule

➝ intracellular trafficking

37
Q

What does HIV use to get entry into the nucleus?

A

➝ NPC

➝ nuclear pore complex

38
Q

What directs the HIV genome into the nucleus?

A

➝ At the NPC it interacts with Nup358 and Nup153

39
Q

What is the protein composition of reverse transcriptase?

A

➝ heterodimers of p66 and p51 subunits

40
Q

In what subunit are the catalytic properties of reverse transcriptase?

A

➝ p66

41
Q

What is the function of p51?

A

➝ structural role and lacks RNAse H domain

42
Q

What are the three enzymatic activities of Reverse transcriptase?

A

1) RNA dependent DNA polymerase
2) RNAse H (cleaves RNA from RNA/DNA hybrid)
3) DNA dependent DNA polymerase

43
Q

What are the three basic steps of reverse transcription?

A

➝ RNA forms intrinsic structures which the enzyme recognises
➝ the RNA primer moves to the other end of the genome
➝ the HIV DNA genome (provirus) is integrated into the host chromosomes

44
Q

How does HIV integrate its genome into the host?

A

➝ There are specific sequences called TTAAs at the end of the HIV genome
➝ the target sequences find each other
➝ the integrase protein recognises the target sequence and flips the integrated form of the genome into the host DNA

45
Q

What is the function of the integrase?

A

➝ loops the viral DNA around the target DNA
➝ it brings the TTAA termus sequences together
➝ it breaks open the host DNA and anneals the viral DNA to the human DNA one strand at a time

46
Q

What does LEDGF/P75 bind to?

A

➝ binds HIV-1 integrase and facilitates targeting to chromatin

47
Q

What is the function of the LEDGF protein?

A

➝ it is bound by the virus and brings it through the nuclear membrane
➝ it allows the integrase protein to recognise target sequences

48
Q

When does the HIV pick up LEDGF?

A

➝ it picks it up on the way to the nucleus

49
Q

What does the HIV-1 promoter contain?

A

➝ binding sites for transcription factors present in T lymphocytes

50
Q

What is transcription controlled by?

What factors promote/ enhance transcription?

A

➝ the binding of transcription factors to the viral DNA

➝ Lef and Nf-Kb - major ones

51
Q

How does the HIV-1 provirus generate different mRNAS for the different viral proteins?

A

➝ splicing

52
Q

What does TAR RNA bind to?

A

➝ The Tat protein

53
Q

What does the TAT-TAR association/binding do?

A

➝ preferentially brings the RNA polymerase II to the genome

➝ enhances elongation of RNA pol II

54
Q

What protein mediates nuclear export of viral RNA?

A

➝ HIV-1 Rev

55
Q

What does the HIV-1 Rev protein mediate?

A

➝ nuclear export of unspliced and singly spliced viral RNA

56
Q

What is HIV Rev essential for?

A

➝ nuclear export of intron containing viral mRNAs

57
Q

What does the HIV Rev protein interact with?

A

➝ HIV-1 Rev protein interacts with Crm1 and RRE RNA

58
Q

What is the mRNA for Gag and Gag-Pol proteins?

A

➝ unspliced HIV-1 RNA

59
Q

What allows packaging of two genomes in HIV?

A

➝ Dimerisation of the unspliced viral RNA

60
Q

How are two genomes able to be packed in HIV?

A

➝Unspliced HIV-1RNA is the mRNA for Gag and Gag-Pol proteins
➝There is an interaction of two RNA genomes (the Gag and Gag-Pol) surrounded by protein
➝In the viral RNA there are kissing loop complexes of RNA that can interact with each other
➝In the SL1 and SL4 domains the loop occurs
➝Dimerisation of the unspliced viral RNA allows packing of two genomes

61
Q

How does HIV get different proteins from the same mRNA?

A

➝ the ‘slippery sequence’
➝ if you start at one end of the genomic RNA you get Gag
➝ if you ‘slip’ the ribosome and get it into a different reading frame you get the Pol protein

62
Q

What generates Gag-pol protein?

A

➝-1 ribosomal frameshifting induced by a ‘slippery’ sequence
➝an RNA hairpin structure

63
Q

How do the proteins get to the plasma membrane?

A

➝ Myristoylation of glycines in the MA domain of Gag mediates association with the plasma membrane

64
Q

What has to be added to the end of the protein so it can associate with the plasma membrane?

A

➝ Myristic acid

65
Q

What is required for virus budding and what does it mediate?

A

➝ HIV-1 PT(S)AP motif

➝ mediates binding of the host Tsg101 protein

66
Q

How does the capsid formation occur?

A

➝P6 binds the Tsg101 protein
➝The ESCRT machinery is hijacked by HIV to exit the membrane
➝During cell division cytokinesis uses the ESCRT machinery
➝The virus hijacks the ESCRT machinery
➝Protease releases the individual proteins from Gag and Gag-Pol polyproteins
➝The polyproteins come together and form the capsid