Week_8_2009_DNADeaminase_Answers

Week_8_2009_DNADeaminase_Answers - Week 8 Make-Up...

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Week 8 Make-Up Assignment Write a 1-2 page summary of the DNA Deamination paper. In your summary, explain the evidences that support the hypothesis they are testing in this paper, then describe the virus reporter system in this study, and what data they obtained to support their hypothesis. Finally, write a concise but complete explanation of the data and state the conclusions made from the data. Can you use the conclusion from this study to help with designing better therapeutic method for retrovrius (esp. HIV) infection? Please explain why.
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DNA Deamination Mediates Innate Immunity to Retroviral Infection Reuben S. Harris, Kate N. Bishop, Ann M. Sheehy, Heather M. Craig, Svend K. Petersen-Mahrt, Ian N. Watt, Michael S. Neuberger, and Michael H. Malim Cell 113: 803-809 (2003) [email protected]
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C4 Deamination Occurs in both RNA (known as “C- to-U editing”) and DNA (known as DNA mutating). Apobec3G is expressed in human T cells APOBEC3G is a potent inhibitor of HIV replication ( Vif -deficient). HIV Vif Vif = viral infectivity factor, one of HIV’s “accessory” gene products viral infectivity, virion assembly, and DNA synthesis necessary in the virus- producing cell for antagonizing the activity of Apobec3G that is co-packed with the progeny virus. APOBECs C U Background Introduction
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Apobec3G: A DNA Mutator CEM15/Apobec3G is a member of the APOBEC family Prototype protein of this family is Apobec1: the catalytic component of the apolipoprotein B editing complex which specifically deaminates C6666 to U in the mRNA of apolipoprotein B Apobec3G is reported as a DNA mutator in E. coli (dC-to-dU mutations) Apobec3G: A DNA Mutator Affecting Virus Infectivity? 1. Prototype protein Apobec1 of APOBEC family edits RNA C-to-U 2. Apobec3G shares concensus sequence required for catalytic function, and is a DNA mutator in E.coli 3. Apobec3G is a potent Vif-deficient HIV inhibitor 4. HIV Vif is indispensable for prodcutive infection by HIV 5. Retrovirus makes DNA molecules
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Model for DNA deamination-mediated anti-retroviral immunity
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(Adapted from Chiu and Greene, Trends Immunol. 2006) Producer Cell Hypothesis: “APOBEC3G triggers the block to retroviral infectivity by deaminating dC dU in the first (minus)-strand of the viral cDNA during a postentry step of its life cycle.” Target Cell
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Question #1: What was the hypothesis (research question) tested in this paper?
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