Intracellular diversion of glycoprotein GP160 of human immunodeficiency virus to lysosomes as a strategy of AIDS gene therapy.
A potential gene therapy strategy against human immunodeficiency virus (HIV-1) is to disrupt the intracellular transport of viral proteins. We report here the binding and transporting of HIV-1 glycoprotein gp160 to lysosomes as a result of the expression of fusion genes consisting of soluble CD4 and lysosome targeting domains. The effective lysosome targeting domain tested includes a lysosomal protease zymogen, procathepsin D, and the COOH-terminal domains of three lysosome membrane proteins: lamp-1, lamp-2, and lysosomal acid phosphatase. We demonstrated that cell fusion (syncytium), caused by the transport of gp160 to the surface of HeLa-CD4+ cells, was completely abolished by the expression of these fusion genes. The lysosomal localization of gp160 in HeLa cells coexpressing CD4-fusion genes was also established. From pulse-chase experiments, we observed that gp160 and the fusion proteins were degraded, as expected of lysosomal activities. Additionally, T lymphoblastoid cells transiently and permanently expressing these fusion genes strongly retarded the propagation of human immunodeficiency virus type 1. Thus, these fusion genes can deprive HIV of newly synthesized envelope protein gp160 for the assembly of new virions and are potentially useful in gene therapy against AIDS.
Lin
X
X
Protein Studies Program, Oklahoma Medical Research Foundation, Oklahoma City 73104.
Dashti
A
A
Schinazi
R F
RF
Tang
J
J
D000135:Acid Phosphatase / Q000235:genetics; D000163:Acquired Immunodeficiency Syndrome / Q000628:therapy*; D001483:Base Sequence; D015704:CD4 Antigens / Q000235:genetics*; D002402:Cathepsin D / Q000235:genetics; D004792:Enzyme Precursors / Q000235:genetics; D015870:Gene Expression; D015686:Gene Products, env / Q000032:analysis / Q000378:metabolism*; D015316:Genetic Therapy / Q000379:methods*; D019302:HIV Envelope Protein gp160; D015497:HIV-1 / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008247:Lysosomes / Q000378:metabolism*; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000032:analysis / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis
D015704:CD4 Antigens; D004792:Enzyme Precursors; D015686:Gene Products, env; D019302:HIV Envelope Protein gp160; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D000135:Acid Phosphatase; C058345:procathepsin D; D002402:Cathepsin D