Formation of noninfectious HIV-1 virus particles lacking a full-length envelope protein.
Deletions were constructed within a functional human immunodeficiency virus type 1 (HIV-1) proviral clone in order to assess the role of the envelope protein in virus particle formation. A graded exonuclease deletion technique was used to produce 12 clones with deletions of 175-308 nucleotides in the first conserved domain of envelope. This included 9 clones with frameshift deletions and 3 clones with in-frame deletions. Isogenic pairs of env deletion clones were produced with or without an additional deletion in the vif and vpr genes. Upon transfection, all clones produced virus particles, as determined by p24 antigen, reverse transcriptase, and sucrose gradient assays with conditioned media. Virus particles produced from clones with deletions in env or vif and vpr, or both regions, banded on sucrose gradients with a mobility similar to that of virus produced by the parental clone. The p24 gag capsid protein in the particles was resistant to trypsin, but the particles were disrupted by treatment with Triton X-100, suggesting the presence of a surrounding lipid bilayer. Furthermore, electron microscopic studies revealed both mature and immature virus particles derived from COS cells transfected with the env deletion clones. Cocultivation experiments with lymphoid cells and cells transfected with each of the env deletion clones demonstrated that the virus particles were noninfectious.
Ratner
L
L
Department of Medicine, Washington University, St. Louis, MO 63110.
Vander Heyden
N
N
Garcia
J
J
Polinsky
M
M
Westervelt
P
P
Becich
M
M
D000818:Animals; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D015683:Gene Products, gag / Q000032:analysis; D015752:Genes, env; D016341:Genes, vif; D016334:Genes, vpr; D015488:HIV Antigens / Q000032:analysis; D016655:HIV Core Protein p24; D015497:HIV-1 / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D008854:Microscopy, Electron; D011533:Proviruses / Q000235:genetics; D012191:Retroviridae Proteins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014758:Viral Core Proteins / Q000032:analysis; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D015683:Gene Products, gag; D015488:HIV Antigens; D016655:HIV Core Protein p24; D012191:Retroviridae Proteins; D014758:Viral Core Proteins; D014759:Viral Envelope Proteins