Fully functional, naturally occurring and C-terminally truncated variant human immunodeficiency virus (HIV) Vif does not bind to HIV Gag but influences intermediate filament structure.
A variant human immunodeficiency virus type 1 (HIV-1) vif gene, vifA45-2, which encodes a protein lacking 19 amino acids at the C terminus but which is fully functional in supporting HIV replication in non-permissive cells has been described previously. By employing newly generated anti-VifA45 serum, further properties of VifA45 and its full-length counterpart, VifA45open, in comparison to Vif from HIV strain BH10 are reported in permissive HeLa and COS-7 cells. The results obtained using confocal microscopic localization studies and in vitro binding assays do not support a requirement for the direct interaction of HIV Gag with Vif. Furthermore and in contrast to previous conclusions, detergent solubility analyses do not demonstrate a role for the C terminus of Vif in mediating localization to the fraction containing cellular membrane proteins. Localization of Vif from HIV strain BH10 to perinuclear aggregates in a small fraction (about 10%) of transfected HeLa cells has been previously reported. The intermediate filament protein vimentin colocalizes to these structures. In contrast, VifA45 and VifA45open form perinuclear aggregates in nearly all transfected HeLa cells; vimentin as well as the cytoskeletal-bridging protein plectin, but not the microtubular protein tubulin, become relocalized to these structures. Interestingly, in COS-7 cells, all of the functional Vif proteins tested (Vif from strain BH10, VifA45 and VifA45open) predominantly localize in the cytoplasm but still induce dramatic aggregation of vimentin and plectin, i.e. in these cells the respective Vif proteins are influencing intermediate filament structure in the absence of colocalization.
Henzler
Tanja
T
Forschungsschwerpunkt Angewandte Tumorvirologie, F02001, and Forschungsschwerpunkt Krebsentstehung und Differenzierung, A01002, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 242, D-69120 Heidelberg, Germany.
Harmache
Abdallah
A
Forschungsschwerpunkt Angewandte Tumorvirologie, F02001, and Forschungsschwerpunkt Krebsentstehung und Differenzierung, A01002, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 242, D-69120 Heidelberg, Germany.
Herrmann
Harald
H
Forschungsschwerpunkt Angewandte Tumorvirologie, F02001, and Forschungsschwerpunkt Krebsentstehung und Differenzierung, A01002, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 242, D-69120 Heidelberg, Germany.
Spring
Herbert
H
Forschungsschwerpunkt Angewandte Tumorvirologie, F02001, and Forschungsschwerpunkt Krebsentstehung und Differenzierung, A01002, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 242, D-69120 Heidelberg, Germany.
Suzan
Marie
M
Pathogénie des Infections à Lentivirus, INSERM U372, BP178, 13276 Marseille, France3.
Audoly
Gilles
G
Pathogénie des Infections à Lentivirus, INSERM U372, BP178, 13276 Marseille, France3.
Panek
Therese
T
Forschungsschwerpunkt Angewandte Tumorvirologie, F02001, and Forschungsschwerpunkt Krebsentstehung und Differenzierung, A01002, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 242, D-69120 Heidelberg, Germany.
Bosch
Valerie
V
Forschungsschwerpunkt Angewandte Tumorvirologie, F02001, and Forschungsschwerpunkt Krebsentstehung und Differenzierung, A01002, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 242, D-69120 Heidelberg, Germany.
D000818:Animals; D019556:COS Cells; D002461:Cell Line, Transformed; D002522:Chlorocebus aethiops; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism*; D016346:Gene Products, vif / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007381:Intermediate Filament Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D051190:Plectin; D011485:Protein Binding; D011817:Rabbits; D012995:Solubility; D014162:Transfection; D014404:Tubulin / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism; D014771:Virion / Q000502:physiology; D054320:vif Gene Products, Human Immunodeficiency Virus
D015683:Gene Products, gag; D016346:Gene Products, vif; D007381:Intermediate Filament Proteins; C495332:PLEC protein, human; D051190:Plectin; D014404:Tubulin; D014746:Vimentin; D054320:vif Gene Products, Human Immunodeficiency Virus