Student Theses and Dissertations

Yang Luo

Date of Award

2007

Document Type

Thesis

Degree Name

Doctor of Philosophy (PhD)

RU Laboratory

Muesing Laboratory

Abstract

Given the limited genetic coding capacity of HIV-1, it is reasonable to expect that the virus must interact with an extensive set of cellular factors and their complexes to complete its passage through the cell. Indeed, it is remarkable that the viral genome, comprising only about 0.0003% of the entire genetic capacity of the cell, commandeers the cellular environment to its own advantage. However, to date, only a small group of cellular proteins have been shown to be required for viral propagation. In an effort to recover and identify those host proteins that interact in complex with the viral machinery, we have developed a systematic genetic method to select derivatives that can encode a small, but potent, foreign epitope tag yet remain fully replication-competent in culture. In conjunction with a novel cryogenic methodology to capture and preserve viralhost interactions usually lost when more conventional isolation techniques are employed, we have recovered host complexes that interact specifically with each of three independently tagged HIV-1 proteins during progressive infection. Thus, the quantitative purification of the tagged viral proteins has allowed the identification of both described factors already known to interact with each of the targeted viral proteins and as well, unanticipated sets of new host proteins in complex with the virus and previously obscured from investigation. Identification and characterization of protein-protein interactions between the host and the virus will provide insight into the cellular processes expropriated by the virus to complete its life cycle.

Comments

A Thesis Presented to the Faculty of The Rockefeller University in Partial Fulfillment of the Requirements for the degree of Doctor of Philosophy

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