Rab27a controls HIV-1 assembly by regulating plasma membrane levels of phosphatidylinositol 4,5-bisphosphate.
During the late stages of the HIV-1 replication cycle, the viral polyprotein Pr55(Gag) is recruited to the plasma membrane (PM), where it binds phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and directs HIV-1 assembly. We show that Rab27a controls the trafficking of late endosomes carrying phosphatidylinositol 4-kinase type 2 α (PI4KIIα) toward the PM of CD4(+) T cells. Hence, Rab27a promotes high levels of PM phosphatidylinositol 4-phosphate and the localized production of PI(4,5)P2, therefore controlling Pr55(Gag) membrane association. Rab27a also controls PI(4,5)P2 levels at the virus-containing compartments of macrophages. By screening Rab27a effectors, we identified that Slp2a, Slp3, and Slac2b are required for the association of Pr55(Gag) with the PM and that Slp2a cooperates with Rab27a in the recruitment of PI4KIIα to the PM. We conclude that by directing the trafficking of PI4KIIα-positive endosomes toward the PM, Rab27a controls PI(4,5)P2 production and, consequently, HIV-1 replication.
Instituto de Investigaciones Biomédicas en Retrovirus y Síndrome de Inmunodeficiencia Adquirida (INBIRS)-Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad de Buenos Aires, C1121ABG Buenos Aires, Argentina.
Pehuén Pereyra
PP
Gerber
Instituto de Investigaciones Biomédicas en Retrovirus y Síndrome de Inmunodeficiencia Adquirida (INBIRS)-Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad de Buenos Aires, C1121ABG Buenos Aires, Argentina.
Mercedes
M
Cabrini
Instituto de Medicina Experimental-CONICET, Academia Nacional de Medicina, C1425AUM Buenos Aires, Argentina.
Carolina
C
Jancic
Instituto de Biologia Molecular y Celular de Rosario-CONICET, S2000EZP Santa Fe, Argentina.
Luciana
L
Paoletti
Instituto de Biologia Molecular y Celular de Rosario-CONICET, S2000EZP Santa Fe, Argentina.
Claudia
C
Banchio
Instituto de Física de Buenos Aires-CONICET, Departamento de Física; and Centro de Microscopías Avanzadas; Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, C1428EGA Buenos Aires, Argentina.
Catalina
C
von Bilderling
Instituto de Física de Buenos Aires-CONICET, Departamento de Física; and Centro de Microscopías Avanzadas; Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, C1428EGA Buenos Aires, Argentina.
Lorena
L
Sigaut
Instituto de Física de Buenos Aires-CONICET, Departamento de Física; and Centro de Microscopías Avanzadas; Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, C1428EGA Buenos Aires, Argentina.
Lía I
LI
Pietrasanta
Instituto de Investigaciones Biomédicas en Retrovirus y Síndrome de Inmunodeficiencia Adquirida (INBIRS)-Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad de Buenos Aires, C1121ABG Buenos Aires, Argentina.
Gabriel
G
Duette
Virus-Cell Interaction Section, HIV Drug Resistance Program, National Cancer Institute, Frederick, MD 21702.
Eric O
EO
Freed
Institut National de la Santé et de la Recherche Médicale U768 and Institut Imagine, Université Paris Descartes-Sorbonne Paris Cité, 75015 Paris, France Institut National de la Santé et de la Recherche Médicale U768 and Institut Imagine, Université Paris Descartes-Sorbonne Paris Cité, 75015 Paris, France.
Genevieve de Saint
Gde S
Basile
Innate Immunity and Inflammation Laboratory, Instituto Gulbenkian de Ciencia, 2780-156 Oeiras, Portugal.
Catarina Ferreira
CF
Moita
Innate Immunity and Inflammation Laboratory, Instituto Gulbenkian de Ciencia, 2780-156 Oeiras, Portugal.
Luis Ferreira
LF
Moita
Centre de Recherche, Institut National de la Santé et de la Recherche Médicale U932, Institut Curie, 75248 Paris, France.
Sebastian
S
Amigorena
Centre de Recherche, Institut National de la Santé et de la Recherche Médicale U932, Institut Curie, 75248 Paris, France.
Philippe
P
Benaroch
Instituto de Investigaciones Biomédicas en Retrovirus y Síndrome de Inmunodeficiencia Adquirida (INBIRS)-Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad de Buenos Aires, C1121ABG Buenos Aires, Argentina.
Jorge
J
Geffner
Instituto de Investigaciones Biomédicas en Retrovirus y Síndrome de Inmunodeficiencia Adquirida (INBIRS)-Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad de Buenos Aires, C1121ABG Buenos Aires, Argentina maostro@fmed.uba.ar.
Matías
M
Ostrowski
D001693:Biological Transport, Active / Q000235:genetics; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D011992:Endosomes / Q000235:genetics / Q000378:metabolism / Q000821:virology; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D019169:Jurkat Cells; D008264:Macrophages / Q000378:metabolism / Q000821:virology; D008565:Membrane Proteins / Q000378:metabolism; D015778:Minor Histocompatibility Antigens; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000235:genetics / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism; D019065:Virus Assembly / Q000502:physiology*; D014779:Virus Replication / Q000502:physiology*; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D000075745:rab27 GTP-Binding Proteins
D008565:Membrane Proteins; D015778:Minor Histocompatibility Antigens; D019269:Phosphatidylinositol 4,5-Diphosphate; C486043:SYTL2 protein, human; D054301:gag Gene Products, Human Immunodeficiency Virus; D000075745:rab27 GTP-Binding Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); C402182:phosphatidylinositol phosphate 4-kinase; C066694:RAB27A protein, human; D020691:rab GTP-Binding Proteins