Date of Award

Spring 1995

Document Type


Degree Name

Master of Science (MS)


Chemistry & Biochemistry



Committee Director

Laura K. Moen

Committee Member

Frank E. Scully, Jr.

Committee Member

Mark S. Elliott

Committee Member

Patricia A. Pleban

Call Number for Print

Special Collections LD4331.C45 S75


The Human Immunodeficiency Virus, type 1 (HIV-1) is the causative agent of the Acquired immunodeficiency Syndrome (AIDS). Currently, AIDS is the leading cause of death amongst young people in the United States. The large increase of HIV-1 infections over the past decade has spawned considerable research in fighting the spread of this disease. One area of intense HIV-1 research has focused on the enzyme Reverse Transcriptase (RT) which is an essential enzyme involved in the replication and life cycle of HIV-1. HIV-1 RT is composed of p66 and p51 subunits and is only active as a heterodimer in vivo. A homodimer composed of p66 subunits has also been found to be active in vitro. Since dimerization is essential for activity, therein exists a potential chemotherapeutic method of RT inhibition.

This study focused on the kinetic parameters of recombinant mutant L289P of HIV-1 RT which has been shown to decrease the association of the homodimer composed of p66 subunits. The region under investigation is thought to be involved in protein-protein interactions. Site directed mutagenesis was previously performed in this laboratory which substituted a proline residue at position 289 for the wild-type leucine residue. The kinetic parameters KM, V max and primer extension reactions were investigated to give insight to the mechanism of this recombinant mutant.

The results indicated that the KM and V max were not significantly different than results previously published for wild-type enzyme. The primer extension reactions also indicated similar pausing sites when compared to wild-type enzyme. These results lead to the conclusion that even though there may be a slight decrease in association of subunits for this mutant, the mutation does not significantly affect template-primer or nucleotide binding to the enzyme and does not affect the inherent pausing mechanism of this enzyme.


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