Effect of mutations within the BPV-1 E1 DNA binding domain on E1-E2 interaction ad the identification and characterization of E1 interacting peptides /
Interaction of papillomavirus proteins E1 and E2 is critical for replication. In an effort to characterize this important E1-E2 interaction we used a yeast two-hybrid system to determine which domains of each protein were important. This study revealed both an N-terminal E1 (E1N)-E2 and a C-termin...
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| Format: | Thesis Book |
| Language: | English |
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[Place of publication not identified] :
[publisher not identified] ;
2001.
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| Subjects: | |
| Online Access: | http://proxy.library.tamu.edu/login?url=http://proquest.umi.com/pqdweb?did=728909651&sid=1&Fmt=2&clientId=2945&RQT=309&VName=PQD |
| Summary: | Interaction of papillomavirus proteins E1 and E2 is critical for replication. In an effort to characterize this important E1-E2 interaction we used a yeast two-hybrid system to determine which domains of each protein were important. This study revealed both an N-terminal E1 (E1N)-E2 and a C-terminal E1 (E1C)-E2 interaction. Upon further mapping of the E1N-E2 interaction, we found that two separate E2 domains, E2TAD (aa 1-194) and E2DBD (aa 326-410) interacted with E1N. The E1 DNA binding domain (E1DBD), which is found within the E1N domain, contains three hydrophilic regions, HR1, HR2, and HR3. As a means of characterizing the E1N-E2 interaction, we tested HR1, HR2, and HR3 E1DBD mutants for their ability to interact with E2TAD and E2DBD, and to function in a transient replication assay. These data revealed that the HR1 and HR3 regions are important in E2TAD interaction, but a lack of E2TAD interaction did not effect viral replication. Only one E1DBD mutant was found to be defective for E2DBD interaction, and consequently was replication defective. This study also identified a hydrophobic region at the C-terminus of the E1DBD, which appears to play a role in replication. HPVs have been strongly linked to cervical cancer. One method of treating this disease would be to inhibit viral replication. Recently, a concept for inhibition of replication through peptide interaction has been developed. If a peptide can be identified which interacts with E1 and inhibits one of E1's many functions, it may ultimately affect the ability of E1 to act as the initiator of viral replication, thus inhibiting the ability of the virus to maintain itself within the host cells. In this study we screened a random peptide library and isolated over one hundred E1 interacting peptides. Thirty-two peptides were sequenced and only one peptide was found to be similar to a known E1 interacting protein. All of the peptides assayed interacted with E1N, while no peptide was found to interact solely with E1C. Finally, in an electromobility shift assay no inhibition of E1-DNA or E1-E2 interaction was found in the presence of any of the four peptides tested. |
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| Item Description: | Vita. "Major Subject: Medical Sciences". |
| Physical Description: | x, 113 leaves : illustrations ; 28 cm. Issued also on microfiche from University Microfilm Inc. |
| Bibliography: | Includes bibliographical references (leaves 95-112). |