A large deletion in the matrix domain of the human immunodeficiency virus gag gene redirects virus particle assembly from the plasma membrane to the endoplasmic reticulum.
Morphogenesis of retroviruses involves assembly of the structural Gag and Gag-Pol polyproteins with subsequent budding of the virus particle from the plasma membrane and proteolytic cleavage by the viral proteinase. The matrix (MA) domain, representing the N-terminal segment of Gag, plays a critical role in this process. We constructed an in-frame deletion in the MA coding region (lacking codons 16 to 99) of the human immunodeficiency virus (HIV) type 1 gag gene. Following transient transfection of the complete proviral DNA carrying the deletion, the mutant polyprotein was synthesized and proteolytically processed like the wild-type polyprotein. However, release of virus particles was reduced approximately 10-fold. The extracellular particles that were released did not contain viral glycoproteins and were noninfectious. Electron micrographs revealed budding of virus particles into the endoplasmic reticulum (ER) of transfected cells and large numbers of particles within the ER. These particles were all immature and morphologically indistinguishable from intracisternal A-type particles, a class of murine endogenous retrovirus elements. Budding structures at the plasma membrane were rarely seen and only a few extracellular particles were observed, but in contrast to those in the ER, these particles had the morphology of mature particles, similar to that of wild-type HIV, except for the lack of surface projections.
Fäcke
M
M
Angewandte Tumorvirologie, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Janetzko
A
A
Shoeman
R L
RL
Kräusslich
H G
HG
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000382:microbiology* / Q000648:ultrastructure; D002522:Chlorocebus aethiops; D004721:Endoplasmic Reticulum / Q000382:microbiology* / Q000648:ultrastructure; D004797:Enzyme-Linked Immunosorbent Assay; D015685:Fusion Proteins, gag-pol / Q000096:biosynthesis; D015870:Gene Expression; D015683:Gene Products, gag / Q000096:biosynthesis / Q000302:isolation & purification; D015750:Genes, gag; D016679:Genome, Viral; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure; D006801:Humans; D007668:Kidney; D007700:Kinetics; D008954:Models, Biological; D009024:Morphogenesis; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D017384:Sequence Deletion; D013997:Time Factors; D014779:Virus Replication
D015685:Fusion Proteins, gag-pol; D015683:Gene Products, gag; D012333:RNA, Messenger