Human Immunodeficiency Virus, type 1 (HIV-1), is the causative agent of the Acquired Immunodeficiency Syndrome (AIDS). HIV-1 reverse transcriptase (RT), a heterodimer p66/p51, has been the major target for treatment of AIDS. The significance of the p51 subunit and the RNase H domain of p66 in terms of their influence on the RNA-dependent DNA synthesis was investigated. Clones of the wildtype HIV-1 RT subunits, p66 and p51, and a recombinant C-terminal deletion mutant, p64, [Barr, P. J. (1987) Bio/Technoloav 5, 486-489] were employed to study the structure-substrate binding relationships of HIV-1 RT. The activity assays of RNA-dependent DNA synthesis on both poly(rA)(dT) and a random base RNA template hybridized with a DNA oligomer showed that p51 significantly affects the enzyme activity. The increase in processivity by p51 in the p66/p51 heterodimer was also demonstrated. These observations suggested that the integrity of p51 is important in subunit-interactions for maintaining a favorable conformation of the enzyme for optimal function. C-terminal deletion in p66 was seen to decrease the processivity. The dissociation constant (Kd) for poly(rA)(dT) obtained by nitrocellulose binding assays suggested that the processivity of HIV
1 RT on poly(rA)(dT) correlated with the affinity for the substrate. The processivity of RT on RNA335-DNA20 was seen to be affected by the pause sites observed on the autoradiograms. The pauses of DNA synthesis tended to occur at positions of template containing poly G-C sequences. The order of processivity observed on RNA335-DNA20 was p64/p64, p66/p66 < p64/p51 < p66/p51. The C-terminal deletion in p66 was shown to affect the ability to extend the DNA strand on RNA template. In those non-wildtype forms of HIV-1 RT (p66/p66, p64/p64, and p64/p51), the affinity for primer-template seemed to be sensitive to the structure of the RNA template as seen when comparing Kds between poly(rA)(dT) and RNA335-DNA20. The wildtype enzyme, p66/p51, appeared to have a similar affinity for both substrates.
","abstract_html":"<p>Human Immunodeficiency Virus, type 1 (HIV-1), is the causative agent of the Acquired Immunodeficiency Syndrome (AIDS). HIV-1 reverse transcriptase (RT), a heterodimer p66/p51, has been the major target for treatment of AIDS. The significance of the p51 subunit and the RNase H domain of p66 in terms of their influence on the RNA-dependent DNA synthesis was investigated. Clones of the wildtype HIV-1 RT subunits, p66 and p51, and a recombinant C-terminal deletion mutant, p64, [Barr, P. J. (1987) Bio/Technoloav 5, 486-489] were employed to study the structure-substrate binding relationships of HIV-1 RT. The activity assays of RNA-dependent DNA synthesis on both poly(rA)(dT) and a random base RNA template hybridized with a DNA oligomer showed that p51 significantly affects the enzyme activity. The increase in processivity by p51 in the p66/p51 heterodimer was also demonstrated. These observations suggested that the integrity of p51 is important in subunit-interactions for maintaining a favorable conformation of the enzyme for optimal function. C-terminal deletion in p66 was seen to decrease the processivity. The dissociation constant (Kd) for poly(rA)(dT) obtained by nitrocellulose binding assays suggested that the processivity of HIV</p> <p>1 RT on poly(rA)(dT) correlated with the affinity for the substrate. The processivity of RT on RNA335-DNA20 was seen to be affected by the pause sites observed on the autoradiograms. The pauses of DNA synthesis tended to occur at positions of template containing poly G-C sequences. The order of processivity observed on RNA335-DNA20 was p64/p64, p66/p66 < p64/p51 < p66/p51. The C-terminal deletion in p66 was shown to affect the ability to extend the DNA strand on RNA template. In those non-wildtype forms of HIV-1 RT (p66/p66, p64/p64, and p64/p51), the affinity for primer-template seemed to be sensitive to the structure of the RNA template as seen when comparing Kds between poly(rA)(dT) and RNA335-DNA20. The wildtype enzyme, p66/p51, appeared to have a similar affinity for both substrates.</p>","abstract_has_math":false,"creators":["Huang, Steve Chien-Wen"],"institution":null,"degree_name":"Doctor of Philosophy (PhD)","degree_level":"Dissertation","degree_discipline":"Chemistry and Biochemistry","degree_department":null,"school":null,"contributors":["Laura K. Moen","Frank Castora","Christopher Osgood","Mark S. Elliot"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":1994,"date_issued":"1994-07-01T07:00:00Z","date_published":"1994-07-01T07:00:00Z","updated_at":"2026-07-24T03:35:08Z","subjects":["HIV-1","AIDS","Reverse transcriptase","Biochemistry","Molecular Biology"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.odu.edu/biomedicalsciences_etds/117","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Laura K. Moen","Frank Castora","Christopher Osgood","Mark S. Elliot"]},{"key":"dc:creator","label":"Author","values":["Huang, Steve Chien-Wen"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2019-09-24T07:00:00Z"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Chemistry and Biochemistry"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Doctor of Philosophy (PhD)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["HIV-1","AIDS","Reverse transcriptase","Biochemistry","Molecular Biology"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://digitalcommons.odu.edu/biomedicalsciences_etds/117"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Human Immunodeficiency Virus, type 1 (HIV-1), is the causative agent of the Acquired Immunodeficiency Syndrome (AIDS). HIV-1 reverse transcriptase (RT), a heterodimer p66/p51, has been the major target for treatment of AIDS. The significance of the p51 subunit and the RNase H domain of p66 in terms of their influence on the RNA-dependent DNA synthesis was investigated. Clones of the wildtype HIV-1 RT subunits, p66 and p51, and a recombinant C-terminal deletion mutant, p64, [Barr, P. J. (1987) Bio/Technoloav 5, 486-489] were employed to study the structure-substrate binding relationships of HIV-1 RT. The activity assays of RNA-dependent DNA synthesis on both poly(rA)(dT) and a random base RNA template hybridized with a DNA oligomer showed that p51 significantly affects the enzyme activity. The increase in processivity by p51 in the p66/p51 heterodimer was also demonstrated. These observations suggested that the integrity of p51 is important in subunit-interactions for maintaining a favorable conformation of the enzyme for optimal function. C-terminal deletion in p66 was seen to decrease the processivity. The dissociation constant (Kd) for poly(rA)(dT) obtained by nitrocellulose binding assays suggested that the processivity of HIV
1 RT on poly(rA)(dT) correlated with the affinity for the substrate. The processivity of RT on RNA335-DNA20 was seen to be affected by the pause sites observed on the autoradiograms. The pauses of DNA synthesis tended to occur at positions of template containing poly G-C sequences. The order of processivity observed on RNA335-DNA20 was p64/p64, p66/p66 < p64/p51 < p66/p51. The C-terminal deletion in p66 was shown to affect the ability to extend the DNA strand on RNA template. In those non-wildtype forms of HIV-1 RT (p66/p66, p64/p64, and p64/p51), the affinity for primer-template seemed to be sensitive to the structure of the RNA template as seen when comparing Kds between poly(rA)(dT) and RNA335-DNA20. The wildtype enzyme, p66/p51, appeared to have a similar affinity for both substrates.
"]},{"key":"dc:title","label":"Title","values":["Structure-Substrate Binding Relationships of HIV-1 Reverse Transcriptase"]}]}],"canonical_facts":{"dc:contributor":["Laura K. Moen","Frank Castora","Christopher Osgood","Mark S. Elliot"],"dc:creator":["Huang, Steve Chien-Wen"],"dc:date.available":["2019-09-24T07:00:00Z"],"dc:description.abstract":["Human Immunodeficiency Virus, type 1 (HIV-1), is the causative agent of the Acquired Immunodeficiency Syndrome (AIDS). HIV-1 reverse transcriptase (RT), a heterodimer p66/p51, has been the major target for treatment of AIDS. The significance of the p51 subunit and the RNase H domain of p66 in terms of their influence on the RNA-dependent DNA synthesis was investigated. Clones of the wildtype HIV-1 RT subunits, p66 and p51, and a recombinant C-terminal deletion mutant, p64, [Barr, P. J. (1987) Bio/Technoloav 5, 486-489] were employed to study the structure-substrate binding relationships of HIV-1 RT. The activity assays of RNA-dependent DNA synthesis on both poly(rA)(dT) and a random base RNA template hybridized with a DNA oligomer showed that p51 significantly affects the enzyme activity. The increase in processivity by p51 in the p66/p51 heterodimer was also demonstrated. These observations suggested that the integrity of p51 is important in subunit-interactions for maintaining a favorable conformation of the enzyme for optimal function. C-terminal deletion in p66 was seen to decrease the processivity. The dissociation constant (Kd) for poly(rA)(dT) obtained by nitrocellulose binding assays suggested that the processivity of HIV
1 RT on poly(rA)(dT) correlated with the affinity for the substrate. The processivity of RT on RNA335-DNA20 was seen to be affected by the pause sites observed on the autoradiograms. The pauses of DNA synthesis tended to occur at positions of template containing poly G-C sequences. The order of processivity observed on RNA335-DNA20 was p64/p64, p66/p66 < p64/p51 < p66/p51. The C-terminal deletion in p66 was shown to affect the ability to extend the DNA strand on RNA template. In those non-wildtype forms of HIV-1 RT (p66/p66, p64/p64, and p64/p51), the affinity for primer-template seemed to be sensitive to the structure of the RNA template as seen when comparing Kds between poly(rA)(dT) and RNA335-DNA20. The wildtype enzyme, p66/p51, appeared to have a similar affinity for both substrates.
"],"dc:identifier":["https://digitalcommons.odu.edu/biomedicalsciences_etds/117"],"dc:subject":["HIV-1","AIDS","Reverse transcriptase","Biochemistry","Molecular Biology"],"dc:title":["Structure-Substrate Binding Relationships of HIV-1 Reverse Transcriptase"],"thesis:degree_discipline":["Chemistry and Biochemistry"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Doctor of Philosophy (PhD)"]},"updated_at":"2026-07-24T03:35:08Z"}