The aphid transmission factor of cauliflower mosaic virus forms a stable complex with microtubules in both insect and plant cells.
We analyzed the distribution of the cauliflower mosaic virus (CaMV) aphid transmission factor (ATF), produced via a baculovirus recombinant, within Sf9 insect cells. Immunogold labeling revealed that the ATF colocalizes with an atypical cytoskeletal network. Detailed observation by electron microscopy demonstrated that this network was composed of microtubules decorated with paracrystalline formations, characteristic of the CaMV ATF. A derivative mutant of the ATF, unable to self-assemble into paracrystals, was also analyzed. This mutant formed a net-like structure, with a mesh of four nanometers, tightly sheathing microtubules. Both the ATF- and the derivative mutant-microtubule complexes were highly stable. They resisted dilution-, cold-, and calcium-induced microtubule disassembly as well as a combination of all three for over 6 hr. CaMV ATF cosedimented with microtubules and, surprisingly, it bound to Taxol-stabilized microtubules at high ionic strength, thus suggesting an atypical interaction when compared with that usually described for microtubule-binding proteins. Using immunofluorescence double labeling we also demonstrated that the CaMV ATF colocalizes with the microtubule network when expressed in plant cells.
Blanc
S
S
Station de Recherches de Pathologie Comparée, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, Christol-les-Alès, France. Blanc@ales.inra.fr
Schmidt
I
I
Vantard
M
M
Scholthof
H B
HB
Kuhl
G
G
Esperandieu
P
P
Cerutti
M
M
Louis
C
C
D000818:Animals; D001665:Binding Sites; D017796:Caulimovirus / Q000502:physiology*; D002460:Cell Line; D002478:Cells, Cultured; D003001:Cloning, Molecular; D007313:Insecta / Q000821:virology*; D008870:Microtubules / Q000821:virology*; D010944:Plants / Q000648:ultrastructure / Q000821:virology*; D016133:Polymerase Chain Reaction; D011523:Protoplasts; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D018411:Spodoptera; D014162:Transfection; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism
D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; C089411:aphid transmission factor, Cauliflower mosaic virus