title: Glutamic residue 438 within the protease-sensitive subdomain of HIV-1 reverse transcriptase is critical for heterodimer processing in viral particles.
abstract: The biological form of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) is a heterodimer consisting of two polypeptides, p66 and p51, which have identical N-termini. The p51 polypeptide is generated by action of viral protease cleaving the p66 polypeptide between residues Phe440 and Tyr441. Dimerization has been mostly studied using bacterially purified RT bearing amino acid changes in either subunit, but not in the context of HIV-1 particles. We introduced changes of conserved amino acid residues 430-438 into the protease-sensitive subdomain of the p66 subunit and analyzed the reverse transcriptase processing and function using purified variants and their corresponding HIV-1 recombinant clones. Our mutational analysis shows that the conserved Glu438 residue is critical for proper heterodimerization and function of virion-associated RT, but not of bacterially expressed RT. In contrast, the conserved Glu430, Glu432, and Pro433 residues are not important for dimerization of virion-associated RT. The network of interactions made by the Glu438 carboxyl group with neighboring residues is critical to protect the Phe440-Tyr441 from cleavage in the context of the p66/p51 heterodimer and may explain why the p66/p51 is not processed further to p51/p51.
authors.affiliation: INSERM U372, 163 avenue de Luminy, BP 178, 13276 Marseille, France.
authors.forename: J M
authors.initials: JM
authors.lastname: Navarro
authors.forename: L
authors.initials: L
authors.lastname: Damier
authors.forename: J
authors.initials: J
authors.lastname: Boretto
authors.forename: S
authors.initials: S
authors.lastname: Priet
authors.forename: B
authors.initials: B
authors.lastname: Canard
authors.forename: G
authors.initials: G
authors.lastname: Quérat
authors.forename: J
authors.initials: J
authors.lastname: Sire
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D019281:Dimerization; D004926:Escherichia coli; D015870:Gene Expression; D018698:Glutamic Acid / Q000235:genetics / Q000378:metabolism*; D016333:HIV Protease / Q000378:metabolism*; D054303:HIV Reverse Transcriptase / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology* / Q000502:physiology; D007700:Kinetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010455:Peptides / Q000235:genetics; D011108:Polymers; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D014771:Virion / Q000502:physiology; D014779:Virus Replication
chemical_list: D010455:Peptides; D011108:Polymers; D018698:Glutamic Acid; D054303:HIV Reverse Transcriptase; D016333:HIV Protease