Dynamic Interaction of HIV-1 Nef with the Clathrin-Mediated Endocytic Pathway at the Plasma Membrane
Article first published online: 26 OCT 2006
Volume 8, Issue 1, pages 61–76, January 2007
How to Cite
Burtey, A., Rappoport, J. Z., Bouchet, J., Basmaciogullari, S., Guatelli, J., Simon, S. M., Benichou, S. and Benmerah, A. (2007), Dynamic Interaction of HIV-1 Nef with the Clathrin-Mediated Endocytic Pathway at the Plasma Membrane. Traffic, 8: 61–76. doi: 10.1111/j.1600-0854.2006.00512.x
- Issue published online: 26 OCT 2006
- Article first published online: 26 OCT 2006
- Received 21 March 2006, revised and accepted for publication 19 October 2006
Figure 1. Nef-DKQTLL mutant colocalizes with clathrin at the plasma membrane. HeLa cells transiently expressing the DKQTLL Nef substitution mutant (A) and DsRed-clathrin (B) were imaged by TIR-FM as in Fig. 2. Insets show higher magnification of a representative area and arrows stress colocalizing spots. Scale bar, 10 μm.
Figure 2. Distribution of the Nef-ERQPLL mutant within dynamic clathrin spot populations. Live HeLa cells expressing either wild-type (black bars) or ERQPLL (grey bars) GFP-tagged Nef fusions in combination with DsRed-clathrin were imaged by TIR-FM. The presence of Nef-GFP inside 40 CS (from more than 5 cells) from each CS population was determined using MetaMorph. Results are expressed as the percentage of clathrin spots of each population that contain Nef (see Fig. 3).
Figure 3: Expression of Nef does not increase the number of clathrin spots at the plasma membrane. HeLa cells transiently expressing wild-type (blue) or LL/AA (white) Nef GFP fusion together with dsRed-clathrin, or CD4-wt (red) or CD4- D cyt (grey) HeLa cell lines expressing Nef-GFP fusions together with DsRed-clathrin were imaged by TIR-FM. For each cell (n=4), the total number of clathrin spots present at the adherent membrane was quantified and then normalized to arbitrary surface unit.
Video 1: Time-lapse movie showing a disappearing spot imaged by TIR-FM which contained both clathrin (top) and Nef (bottom). ~ 0.3 s/frame.
Video 2: Time-lapse movie showing a laterally mobile spot imaged by TIR-FM which contained both clathrin (top) and Nef (bottom). ~ 0.3 s/frame.
|tra_525_sm_8_1cfig1.jpg||495K||Supporting info item|
|tra_525_sm_8_1cfig2.jpg||208K||Supporting info item|
|tra_525_sm_8_1cfig3.jpg||413K||Supporting info item|
|tra_525_sm_8_1cmov1.mpeg||42K||Supporting info item|
|tra_525_sm_8_1cmov2.mpeg||687K||Supporting info item|
Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.