Efficient particle formation can occur if the matrix domain of human immunodeficiency virus type 1 Gag is substituted by a myristylation signal.
Lentiviruses, such as human immunodeficiency virus type 1 (HIV-1), assemble at and bud through the cytoplasmic membrane. Both the matrix (MA) domain of Gag and its amino-terminal myristylation have been implicated in these processes. We have created HIV-1 proviruses lacking the entire matrix domain of gag which either lack or contain an amino-terminal myristate addition sequence at the beginning of the capsid domain. Myristate- and matrix-deficient [myr(-)MA(-)] viruses produced after transient transfection are still able to assemble into particles, although the majority do not form at the plasma membrane or bud efficiently. Myristylation of the amino terminus of the truncated Gag precursor permits a much more efficient release of the mutant virions. While myr(-)MA(-) particles were inefficient in proteolytic processing of the Gag precursor, myristylation enabled efficient proteolysis of the mutant Gag. All matrix-deficient viruses are noninfectious. Particles produced by matrix-deficient mutants contain low levels of glycoproteins, indicating the importance of matrix in either incorporation or stable retention of Env. Since matrix-deficient viruses contain a normal complement of viral genomic RNA, a role for MA in genomic incorporation can be excluded. Contrary to previous reports, the HIV-1 genome does not require sequences between the 5' splice donor site and the gag start codon for efficient packaging.
Lee
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PP
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Linial
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ML
D001483:Base Sequence; D015153:Blotting, Western; D003062:Codon / Q000235:genetics; D015683:Gene Products, gag / Q000378:metabolism*; D016655:HIV Core Protein p24 / Q000032:analysis / Q000096:biosynthesis; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000254:growth & development* / Q000378:metabolism / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D016296:Mutagenesis; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D011499:Protein Processing, Post-Translational; D011533:Proviruses / Q000254:growth & development* / Q000378:metabolism / Q000648:ultrastructure; D012334:RNA, Neoplasm / Q000032:analysis / Q000096:biosynthesis; D012367:RNA, Viral / Q000032:analysis / Q000096:biosynthesis; D012194:RNA-Directed DNA Polymerase / Q000032:analysis / Q000096:biosynthesis; D014162:Transfection; D014763:Viral Matrix Proteins / Q000096:biosynthesis
D003062:Codon; D015683:Gene Products, gag; D016655:HIV Core Protein p24; D009227:Myristic Acids; D009838:Oligodeoxyribonucleotides; D012334:RNA, Neoplasm; D012367:RNA, Viral; D014763:Viral Matrix Proteins; D019814:Myristic Acid; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase