11-16-09 Introduction to DNA Viruses I (aka adenovirus and parvovirus) 1.

Describe the structure of adenovirus and parvovirus Adenovirus structure Naked (no envelope) Medium size Linear double-stranded DNA molecule Important components: CAPSID: penton base, hexon, and fiber CORE: DNA and associated proteins Parvovirus structure Naked (no envelope) icosahedral Small!! Linear single-stranded DNA molecule Important components: NS proteins: non-structural proteins required for DNA replication CAP proteins: structural proteins that comprise the viral capsid 2. Understand the life cycle and mechanism of DNA replication for adenovirus and parvovirus. Adenovirus life cycle Adsorption/attachment is a two-stage process: 1. initial interaction of fiber protein* with a range of cellular receptors such as MHC Class I and the coxackievirus-adenovirus receptor (CAR) 2. the penton base then binds to the integrin family of cell surface receptors Both interactions are required before internalization via receptor-mediated endocytosis is allowed *side note: most cells express primary receptors for fiber proteins, but internalization is more selective Penetration Involves virus phagocytosis Rupture of endosome Pentons have a toxic effect on the endosome leading to its opening, releasing the viral particle into the cytoplasm Uncoating (cytoplasm) Uncoating is a sequential process that actually begins prior to endosomal opening Disassembly of the viral particle continues after release into the cytoplasm Transport into nucleus The viral core migrates into the host cell nucleus DNA enters through the nuclear Gene expression: transcription occurs in three temporally distinct phases: Immediate early genes: Gene transcribed: E1A

Produces regulatory factors that are required for subsequent phases of gene expression + alteration of cellular growth properties that favor virus survival and proliferation Early genes: Genes transcribed: EIB, E2A, E2B, E3, E4, some other virion proteins Encoded at various locations on both strands of DNA; multiple gene products listed above are produced by alternative splicing Produces proteins required for viral DNA replication: DNA polymerase, precursor Terminal protein (pTP), and single strand DNA binding protein (DBP). Regulated by virus encoded trans-acting regulatory factors in coordination with cellular factors, plus viral DNA polymerase II Late genes: Genes transcribed: virion proteins Encodes for structural proteins During and after DNA replication ,most of the viral genes expressed are late genes (makes sense given their function of encoding structural proteins) Genome replication: DNA replication occurs in the nucleus of the host cell 1. viral genome is coated with DBP 2. NFI (nuclear factor I), a cellular protein, binds to its recognition site within the origin of replication. This is followed by the binding of a second host protein, NFIII. There are actually two origins of replication at which this process can occur 3. pTP (precursor terminal protein) and pol (DNA polymerase) are recruited to the initiation complex 4. a covalent link is formed between deoxycytidine triphosphate (dCTP) and a serine residue in pTP. The dCMP-pTP complex serves as a primer for the synthesis of the DNA strand. This is in contrast to the conventional RNA primers Virus assembly 1. begins in the cytoplasm when individual monomers form hexon and penton capsomers. 2. empty, immature capsids are assembled in the nucleus, where the core is formed from genomic DNA + associated core proteins. 3. as virus particles accumulate in the nucleus, they eventually cause cell lysis. They are visible under the microscope as crystals.

Parvovirus life cycle virus binds to erythrocyte P antigen, a glycolipid receptor molecule replication occurs in the nucleus. Important point!!!: parvovirus is highly dependent on cellular functions for genome replication and requires a host cell that passes through S phase for replication to occur

Consequently, parvovirus is only successful in completing life cycles in host cells that proliferate rapidly NS proteins are required for DNA replication CAP proteins are structural proteins that make up the capsid the ends of the parvovirus genome have palindromic sequences (inverted repeats) that allow for the formation of hairpin structures that are essential for the initiation of genome replication the double-stranded version of the viral genome is required for transcription and replication

3. Understand the mechanism of immune evasion by adenovirus. There are two mechanisms of immune evasion by adenovirus inhibition of interferon signaling E1A proteins inhibit signaling pathway Virus-encoded VA RNAs inhibit viral RNA-activated protein kinase PKr. Inhibition of antigen presentation by MHC class I molecules E3 protein prevents antigen presentation by MHC class I E1A protein represses the actual expression of MHC class I genes 4. Describe the diseases associated with adenovirus and parvovirus infection. Understand the concept of biphasic disease in parvovirus. Adenovirus pathogenesis most commonly associated with upper respiratory tract (URT) infections also: pharyngitis, gastroenteritis, conjunctivitis/keratoconjunctivitis (dirty pools), pneumonia, acute hemorrhagic cystitis, hepatitis diseases other than URT infections most commonly infect children and military recruits spread by aerosol, close contact, and oral-fecal contact infects mucoepithelial cells of respiratory tract, GI tract, and cornea; persists in lymphoid tissue Parvovirus pathogenesis a biphasic disease initial phase of viremia: erythrocyte production is halted. Antibodies control the viremia and lead to disease resolution second phase is immune-related. The Abs that stop the initial phase cause the formation of immune complexes that result in rash and arthralgia. Second phase is when specific B19 (Fifth Disease) symptoms (the slapped cheek facial rash) appear and allow for diagnosis In addition to Fifth Disease, parvovirus also causes Intrauterine Fetal Death (due to vertical transmission from mother to fetus during the first trimester; infection and destruction of rbcs) and Polyarthropathy Syndrome (also due to immune complex formation) Check out the table on the last page comparing parvovirus and adenovirus!!