Keywords

HIV, retrocyclin, defensin, innate immunity, docking

Abstract

Retrocyclin RC-101, a θ-defensin with lectin-like properties, potently inhibits infection by many HIV-1 subtypes by binding to the heptad repeat (HR)-2 region of gp41 and preventing six-helix bundle formation. In the present study, we used in silico computational exploration to identify residues of HR2 that interacted with RC-101 and then analyzed the HIV-1 Sequence Database at LANL for residue variations in the HR1 and HR2 segments that could plausibly impart in vivo resistance. Docking RC-101 to gp41 peptides in silico confirmed its strong preference for HR2 over HR1, and implicated residues crucial for its ability to bind HR2. We mutagenized these residues in pseudotyped HIV-1 JR.FL reporter viruses, and subjected them to single round replication assays in the presence of 1.25-10ug/ml RC-101. Except for one mutant that was partially resistant to RC-101, the other pseudotyped viruses with single-site cationic mutations in HR2 manifested absent or impaired infectivity or retained wild-type susceptibility to RC-101. Overall, these data suggest that most mutations capable of rendering HIV-1 resistant to RC-101 will also exert deleterious effects on the ability of HIV-1 to initiate infections - an interesting and novel property for a potential topical microbicide.

Notes

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Graduation Date

2007

Semester

Spring

Advisor

Cole, Alexander

Degree

Master of Science (M.S.)

College

Burnett College of Biomedical Sciences

Department

Molecular Biology and Microbiology

Degree Program

Molecular and Microbiology

Format

application/pdf

Identifier

CFE0001707

URL

http://purl.fcla.edu/fcla/etd/CFE0001707

Language

English

Release Date

June 2007

Length of Campus-only Access

None

Access Status

Masters Thesis (Open Access)

Restricted to the UCF community until June 2007; it will then be open access.

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