title: Preferential recognition of monomeric CCR5 expressed in cultured cells by the HIV-1 envelope glycoprotein gp120 for the entry of R5 HIV-1.
abstract: Bimolecular fluorescence complementation (BiFC) and western blot analysis demonstrated that CCR5 exists as constitutive homo-oligomers, which was further enhanced by its antagonists such as maraviroc (MVC) and TAK-779. Staining by monoclonal antibodies recognizing different epitopes of CCR5 revealed that CCR5 oligomer was structurally different from the monomer. To determine which forms of CCR5 are well recognized by CCR5-using HIV-1 for the entry, BiFC-positive and -negative cell fractions in CD4-positive 293T cells were collected by fluorescent-activated cell sorter, and infected with luciferase-reporter HIV-1 pseudotyped with CCR5-using Envs including R5 and R5X4. R5 and dual-R5 HIV-1 substantially infected BiFC-negative fraction rather than BiFC-positive fraction, indicating the preferential recognition of monomeric CCR5 by R5 and dual-R5 Envs. Although CCR5 antagonists enhanced oligomerization of CCR5, MVC-resistant HIV-1 was found to still recognize both MVC-bound and -unbound forms of monomeric CCR5, suggesting the constrained use of monomeric CCR5 by R5 HIV-1.
authors.affiliation: Department of Medical Virology, Faculty of Life Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Nakano
authors.affiliation: Department of Medical Virology, Faculty of Life Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Monde
authors.affiliation: Department of Medical Virology, Faculty of Life Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Terasawa
authors.affiliation: Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Kami-youga 1-18-1, Setagaya, Tokyo 158-8501, Japan.
authors.forename: Yuzhe
authors.initials: Y
authors.lastname: Yuan
authors.affiliation: Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Kami-youga 1-18-1, Setagaya, Tokyo 158-8501, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Yusa
authors.affiliation: Department of Medical Virology, Faculty of Life Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Harada
authors.affiliation: Department of Medical Virology, Faculty of Life Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan. Electronic address: ymaeda@kumamoto-u.ac.jp.
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Maeda
mesh_terms: D002460:Cell Line; D015699:HIV Envelope Protein gp120 / Q000235:genetics / Q000378:metabolism*; D015658:HIV Infections / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D019713:Receptors, CCR5 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015728:Receptors, HIV / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D053586:Virus Internalization
chemical_list: D015699:HIV Envelope Protein gp120; D019713:Receptors, CCR5; D015728:Receptors, HIV
keywords: Bimolecular fluorescence complementation assay; CCR5; CCR5 antagonist; HIV-1; Monomer; Oligomerization