Analysis of the murine leukemia virus R peptide: delineation of the molecular determinants which are important for its fusion inhibition activity.
In previous studies, the C-terminal R peptide of the murine leukemia virus (MuLV) Env protein was shown to be a potent inhibitor of viral fusion activity. In the present study, we investigated the molecular determinants in the MuLV Env protein cytoplasmic tail which are important for the fusion inhibition activity of the R peptide. We constructed a series of mutant MuLV env genes which express Env proteins with serial truncations, internal deletions, or amino acid substitutions in the cytoplasmic tail. To analyze their cell fusion activity, we employed a quantitative fusion assay. We found that truncations of up to 7 amino acids from the C terminus of the cytoplasmic tail had no detectable effect on the lack of fusion activity of the full-length Env protein; however, further truncations resulted in a progressive increase in cell fusion activity. Studies of mutant proteins with amino acid substitutions in the cytoplasmic tail showed that Leu-627 plays an important role in fusion inhibition by the R peptide, while most of the other amino acids in the R peptide were not essential for fusion inhibition. Studies of mutant proteins with internal deletions upstream of the cleavage site in the cytoplasmic tail showed that this region is also involved in fusion inhibition by the R peptide, although only to a limited extent. The results are consistent with a model in which the MuLV R peptide exhibits its fusion inhibition activity through interaction with a cellular factor(s).
Yang
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Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
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D000595:Amino Acid Sequence; D003593:Cytoplasm / Q000737:chemistry; D015686:Gene Products, env / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D009052:Leukemia Virus, Murine / Q000737:chemistry*; D008561:Membrane Fusion; D008969:Molecular Sequence Data; D010446:Peptide Fragments; D016415:Sequence Alignment; D017384:Sequence Deletion; D013329:Structure-Activity Relationship
D015686:Gene Products, env; D010446:Peptide Fragments