Annexin 2 is not required for human immunodeficiency virus type 1 particle production but plays a cell type-dependent role in regulating infectivity.
During assembly and budding of retroviruses, host cell proteins are incorporated into viral particles. Identification of virion-associated proteins may help pinpoint key cellular components required for virus production and function. The cellular protein annexin 2 (Anx2) is incorporated into HIV-1 particles, and knockdown of Anx2 has been reported to cause defects in Gag processing and infectivity of HIV-1 particles in macrophages. Here, we tested whether Anx2 was required for HIV-1 production in other cell types capable of producing HIV-1 virions. Endogenous Anx2 levels were knocked down by approximately 98% using lentivirus encoding short hairpin RNAs (shRNAs) or small interfering RNAs (siRNAs) targeting Anx2. Under these conditions, there was no reduction in HIV-1 virus-like particle (VLP) production in either COS-1, 293T, or Jurkat T cells or primary human monocyte-derived macrophages (MDMs). Murine embryonic fibroblasts derived from Anx2(-/-) mice produced the same levels of VLPs as matched cells from wild-type mice. The calcium-mediated spike in VLP production still occurred in Anx2-depleted COS-1 cells, and there was no apparent alteration in the intracellular Gag localization. Overexpression of Anx2 in trans had no effect on Gag processing or VLP production. Neither Anx2 depletion nor Anx2 overexpression altered the infectivity of HIV-1 particles produced by COS-1 or 293T cells. However, supernatants containing virus from Anx2 siRNA-treated primary human MDMs exhibited decreased infectivity. These data indicate that Anx2 is not required for HIV-1 assembly or Gag processing but rather plays a cell type-dependent role in regulating production of infectious HIV-1 by macrophages.
Rai
Tia
T
Cell Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, Box 143, New York, NY 10065, USA.
Mosoian
Arevik
A
Resh
Marilyn D
MD
D000818:Animals; D017306:Annexin A2 / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D001483:Base Sequence; D019556:COS Cells; D002460:Cell Line; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D066246:ErbB Receptors / Q000378:metabolism; D055785:Gene Knockdown Techniques; D015497:HIV-1 / Q000472:pathogenicity* / Q000502:physiology*; D054884:Host-Pathogen Interactions / Q000502:physiology; D006801:Humans; D019169:Jurkat Cells; D008264:Macrophages / Q000502:physiology / Q000821:virology; D051379:Mice; D018345:Mice, Knockout; D034741:RNA, Small Interfering / Q000235:genetics; D009418:S100 Proteins / Q000502:physiology; D014771:Virion / Q000502:physiology; D014774:Virulence / Q000502:physiology; D019065:Virus Assembly / Q000502:physiology; D057074:Virus Release / Q000502:physiology; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000378:metabolism; D000099248:S100 Calcium Binding Protein A10
D017306:Annexin A2; D066246:ErbB Receptors; D034741:RNA, Small Interfering; D009418:S100 Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; D000099248:S100 Calcium Binding Protein A10; C472822:ANXA2 protein, human