HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 278, Issue 47, 46601-46606, November 21, 2003

This Article Abstract Full Text (PDF) All Versions of this Article: 278/47/46601    most recent M302221200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Bard, F. Articles by Jurdic, P. Search for Related Content PubMed PubMed Citation Articles by Bard, F. Articles by Jurdic, P. Social Bookmarking                      What's this?

Src Regulates Golgi Structure and KDEL Receptor-dependent Retrograde Transport to the Endoplasmic Reticulum^*

Frédéric Bard, Laetitia Mazelin¶, Christine Péchoux-Longin||, Vivek Malhotra**, and Pierre Jurdic**

From the University of California San Diego Biological Sciences Division, Cell and Developmental Biology Department, University of California San Diego, La Jolla, CA 92093-0347, the ^¶Centre de Génétique Moléculaire et Cellulaire, Unité Mixte de Recherche 5534 CNRS/Université Claude Bernard, Batiment Grégoire Mendel, 16 Rue Dubois, 69622 Villeurbanne Cedex, France, and the ^||Laboratoire de Génomique et Physiologie de la Lactation, Institut National de la Recherche Agronomique, Domaine de Vilvert, 78352 Jouy-en-Josas Cedex and the Laboratoire de Biologie Moléculaire et Cellulaire, Unité Mixte de Recherche 5665 CNRS/ENS, Institut National de la Recherche Agronomique 913, Ecole Normale Supérieure de Lyon, 46, Allée d'Italie, 69007 Lyon, France

Received for publication, March 4, 2003 , and in revised form, August 21, 2003.

	ABSTRACT

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS DISCUSSION REFERENCES

 
The tyrosine kinase Src is present on the Golgi membranes. Its role, however, in the overall function and organization of the Golgi apparatus is unclear. We have found that in a cell line called SYF, which lacks the three ubiquitous Src-like kinases (Src, Yes, and Fyn), the organization of the Golgi apparatus is perturbed. The Golgi apparatus is composed of collapsed stacks and bloated cisternae in these cells. Expression of an activated form of Src relocated the KDEL receptor (KDEL-R) from the Golgi apparatus to the endoplasmic reticulum. Other Golgi-specific marker proteins were not affected under these conditions. Because of the specific effect of Src on the location of KDEL-R, we tested whether protein transport between ER and the Golgi apparatus involves Src. Transport of Pseudomonas exotoxin, which is transported to the ER by binding to the KDEL-R is accelerated by inhibition or genetic ablation of Src. Protein transport from ER to the Golgi apparatus however, is unaffected by Src deletion or inhibition. We propose that Src has an appreciable role in the organization of the Golgi apparatus, which may be linked to its involvement in protein transport from the Golgi apparatus to the endoplasmic reticulum.

	INTRODUCTION

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS DISCUSSION REFERENCES

 
Src is activated by various cell surface receptors and phosphorylates numerous cellular proteins (for review, see Refs. 1 and 2). A significant fraction of Src is localized to the perinuclear region and the secretory vesicles in neuronal cells (3, 4). It interacts with and phosphorylates proteins such as dynamin and synapsin (5–7) and synaptophysin (8) or cellugyrin (9), suggesting a role in the regulation of membrane traffic. Src and other Src family tyrosine kinases are also localized to the Golgi apparatus (3, 10). On the Golgi apparatus, Src interacts with the ubiquitin ligase Cbl, and Src activation leads to an increased recruitment of Cbl specifically at the Golgi. This indicates that Src-Cbl signaling might occur at this organelle (10). Further support for this proposal stems from recent findings that show that Src is involved in the activation of Ras on the Golgi apparatus following mitogenic stimulation (11). Src is also known to phosphorylate and interact with a Golgi apparatus-specific protein, Golgin 67 (12). However, the function of Src signaling at the Golgi remains unknown. The number of signaling molecules that associate with the Golgi membranes and regulate its function and organization is increasing and is known to include protein kinase D, which regulates Golgi-to-plasma membrane trafficking (13) (14), and mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK) and polo-like kinase, which are essential for the fragmentation of the Golgi at the onset of mitosis (15) (16) (17). It is thus reasonable to propose that Src might also be involved in the structural and functional regulation of the Golgi membranes.

A major issue in studies involving Src is the functional redundancy between the Src kinase family, which includes nine members (18). Knock-out studies have revealed that the deletion of Src has relatively mild effects in mice. However, Src knock-out in combination with a deletion of Yes and Fyn, the two other members of the Src family, leads to an early embryonic death (19). A cell line named SYF¹ (for Src, Yes, and Fyn triple knock-out) has been generated from one of these embryos and provides a unique tool to analyze the cellular function of Src.

We found that SYF cells have an aberrant Golgi morphology with compacted stacks and bloated cisternae. Furthermore, expression of an activated form of Src induces the specific exclusion of the KDEL-R from the Golgi apparatus, suggesting an effect on retrograde trafficking. Consistently, the rate of traffic between the Golgi apparatus and the ER of the Pseudomonas exotoxin is reduced by Src expression. Deletion of Src does not appear to perturb protein transport in the anterograde pathway from the ER to the plasma membrane via the Golgi apparatus.

	EXPERIMENTAL PROCEDURES

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS DISCUSSION REFERENCES

 
Reagents and Cells—The polyclonal Src2 and monoclonal B-12 anti-Src antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA), and the KDEL-R antibody (clone KR-10) was from Calbiochem. Rabbit anti-mannosidase II is a generous gift of Dr. Kelly Morgan (University of Georgia). SYF, SYFsrc, and Src+ cells were a kind gift of Drs. Klinghoffer and Soriano (Seattle, WA). Src inhibitor SU6656 was purchased from Calbiochem Novabiochem, and Pseudomonas exotoxin came from List Biological Laboratories (Campbell, CA).

Electron Microscopy—Monolayers of SYF and Src+ cells were washed with PBS and fixed in situ with 2.5% glutaraldehyde in phosphate buffer, pH 7.4, for 30 min at 4 °C. Cells were then postfixed with 1% osmium tetroxide containing 1.5% potassium cyanoferrate, dehydrated in gradual ethanol (70% to 100%), and embedded in Epon. Thin sections were applied onto 200 mesh copper grids and counterstained with uranyl acetate and lead citrate before examination with a Philips CM 120 transmission electron microscope (CMEABG, Villeurbanne, France).

Pseudomonas Exotoxin Transport Assay—SYF and Src+ cells were grown to near confluence in 24-well plates. 1 h prior to and then during the experiment, cells were incubated in Dulbecco's modified Eagle's medium, without methionine/cysteine, containing 25 mM Hepes. The cells were then incubated with 20 µg/ml Pseudomonas exotoxin (PE) or in medium alone (control) for 30 min at 19 °C and then shifted to 37 °C for various periods of time in three wells for PE and three wells of control for each time point. 10 µCi of trans-³⁵S mixture (ICN Pharmaceuticals) was added to each well for 10 min, and the wells were washed twice with PBS before cell lysis and trichloroacetic acid precipitation. After resuspension in sample buffer, the radioactivity in each sample was quantified by scintillation counter, and the average value of treated wells was normalized to the average value of control wells. As an additional control, brefeldin A was added at 5 µg/ml in three wells in the same time as was the PE. For Src drug treatment, Src+ cells were pretreated with 2 µM SU6656 for 30 min prior to the addition of PE.

Vesicular Stomatitis Virus (VSV)-G Protein Transport Assay—GFP-VSV-G protein was transfected using LipofectAMINE® (Invitrogen) according to the manufacturer's recommendation. 24 h after transfection, the cells were transferred at 40 °C for 6 h. Cycloheximide was then added at a concentration of 100 µg/ml. At this stage, which was taken as the 0 min time point, cells were transferred at 32 °C for the indicated period of time before fixation for fluorescence-based analysis.

Immunofluorescence Microscopy—Cells were fixed in 4% formaldehyde in PBS and treated with PBS containing 1% bovine serum albumin and 0.1% Triton X-100 for 1 h before incubation with primary antibodies. Alexa 488- and Alexa 594-conjugated antibodies (Molecular Probes, Eugene, OR) were used as secondary antibodies. Coverslips were mounted with Cytoseal 60 (Richard-Allan Scientific, VWR International) and visualized with a Nikon Microphot-FXA microscope with a Plan-APO 60X objective. Digital images were acquired with an Olympus camera.

Image Analysis and Quantification—The freeware ImageJ 1.29x by Wayne Rasband was used for image quantification and analysis. To quantify the morphology of the Golgi apparatus, the immunofluorescence of mannosidase II was recorded as a digital, 8-bit gray-scale, 1280 x 1024 pixels image. The Golgi labeling image threshold was set at 90 on a 0–255 black to white scale (all pixels with a value under 90 are excluded from the quantification) to remove background pixels from measurement. A region of interest was defined for each cell, and the perimeter and surface of the Golgi apparatus with a minimum area of three pixels was measured. The dimensionless circularity of the Golgi apparatus was then computed according to the formula 4(sum (areas)/(sum(perimeters))^2). To quantify the decrease of the KDEL-R immunofluorescence signal at the Golgi in Src-transfected cells, the 8-bit grayscale, non-saturated images of KDEL-R or mannosidase II (as a control) immunofluorescence were set at a threshold value of 90). For each cell, the number of pixels over threshold and their mean value were measured inside a region of interest covering the perinuclear area. The product of the mean intensity and the number of pixels for each cell was considered the value of immunofluorescence intensity for KDEL-R in the given perinuclear area. Each microscopic field contained one transfected cell and 3–6 control cells, and the average of labeling intensity for the control cell was used to normalize the data. For GFP-VSV-G protein transport assay quantification, we measured the ratio of the average green fluorescence intensity of pixels above background between the Golgi area and the rest of the cell. For each GFP-VSV-G protein green channel image, we used the corresponding GRASP65 (Golgi resident protein) red channel image to generate a "filter," i.e. a binary image in which the value of each pixel below background is set to 0, and the value for each pixel above it is set to 1. Each pixel of this binary image was then multiplied by the corresponding pixel (value 0–255) from the green channel image, and the resulting image was then used to measure the average GFP fluorescence above background per pixel in the Golgi area. An inverted image of the filter (0 replaced by 1 and reciprocally) was used to measure the GFP fluorescence above background in the rest of the cell.

Statistical Analysis—For statistical computation and estimation of significance, we used the online software GraphPad® (www.graphpad.com). The circularity of the Golgi apparatus in multiple cells and the normalized results of KDEL-R fluorescence at the Golgi for control and transfected cells were run through unpaired t tests. According to GraphPad® recommendations, the computed two-tailed p values were considered extremely statistically significant when p < 0.001 (noted as *** in Figs. 1G and 3D), very significant when p ranges from 0.001 to 0.01 (noted as ** in Fig. 1G), significant when 0.05 > p > 0.01, and not significant (n/s in Fig. 3H) when p > 0.05.

View larger version (25K): [in this window] [in a new window]   FIG. 1. SYF cells present a Golgi-compacted morphology. Src+ (A, B, and C) and SYF (D, E, and F) cells were stained with Hoescht (A and D) and a polyclonal anti-mannosidase II (B and E). To quantify the morphology of the Golgi, digital images of mannosidase II labeling were set at a threshold of 90 (C and F), excluding all pixels under the threshold intensity value from measurement. The area and the perimeter of each Golgi element of a particular cell were then computed, and their respective sums were used to calculate the compactness of the organelle. The measures for the images in B and E are shown in C and F (where P is sum of perimeter of Golgi units, A is the sum of the areas of Golgi units, and C is the compactness value derived from P and A. See "Experimental Procedures" for details). The compactness values of 22 SYF, 10 Src+, and 9 SYFsrc cells were computed and submitted to statistical analysis (G). ***, extremely statistically significant p < 0.001; **, very statistically significant p = 0.0014. Bar is 10 µm in B and F.

View larger version (32K): [in this window] [in a new window]   FIG. 3. The KDEL-R is redistributed following SrcE378G transfection. SYF cells were transfected with a SrcE378G expression plasmid, fixed 12 h later, and labeled with the anti-Src polyclonal antibody Src2 (A) and the anti KDEL-R monoclonal antibody (B) or the monoclonal anti phosphotyrosine PY99 (E) and the polyclonal anti-mannosidase II (F). To quantify the fluorescence at the Golgi, the digital images were set at a threshold of 90 (C and G), and the total number of pixels above threshold, their mean intensity, and the fluorescent intensity in the perinuclear area (circles) of each cell were computed. In panels C and G, the particular measurements for the images B and F, respectively, are shown (A denotes the number of pixels, M is the mean value of intensity, and I is fluorescent intensity, the product of A and M). Each intensity value was then normalized to the mean intensity of control cells (CON) in the same image to allow statistical analysis (D and H); 27 control cells and 7 transfected cells were analyzed for KDEL-R, and 28 control cells and 10 transfected cells were analyzed for mannosidase II. Arrows point to transfected cells. ***, extremely statistically significant; n/s, not statistically significant. Bar in panels B and F is 10 µm.

	RESULTS

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS DISCUSSION REFERENCES

To further analyze the organization of collapsed Golgi apparatus, Src+, SYFsrc, and SYF cells were imaged by electron microscopy. 10 cells for each cell line and 4–5 serial sections for each cell were imaged; the most representative photographs are shown in Fig. 2. In SYF cells, the Golgi apparatus was collapsed into a compact structure in which the individual stacks could not be identified (Fig. 2A). Interestingly, the individual Golgi cisternae were enlarged in SYF cells (Fig. 2A, inset) compared with their Src+ counterparts (Fig. 2B, inset). In Src+ (Fig. 2B) and SYFsrc cells (not shown), stacks appeared normal. This overall Golgi organization in SYF cells compares well with the organization of the Golgi apparatus observed at the immunofluorescence level.

View larger version (122K): [in this window] [in a new window]   FIG. 2. The compacted Golgi morphology in SYF cells results from collapsed stacks and bloated cisternae. SYF (A), Src+ (B), and SYFsrc (not shown) cells were treated for electron microscopy as described under "Experimental Procedures." 10 cells of each cell line and 4–5 serial sections for each cell, covering most of the Golgi apparatus, were imaged. Representative micrographs of the two different morphologies observed are shown in panels A and B. Insets in panels A and B are a magnification of the area of the Golgi stacks. Bar represents 0.4 µm.

Src Expression Reduces the Rate of Golgi-to-ER Traffic of the Pseudomonas Exotoxin—The effect of Src on KDEL-R distribution suggests that Src may be involved in retrograde traffic from the Golgi apparatus to the ER. We used the bacterial PE, which is transported after endocytosis to the ER via the Golgi apparatus (21). PE binds to KDEL-R by its KDEL-like sequence that insures its transport to the ER. Upon reaching the ER, it is translocated to the cytosol and inhibits protein synthesis (22). To test the kinetics of transport of PE, we measured the delay between PE internalization and the onset of protein synthesis inhibition induced by PE. SYF, SYFsrc, or SYFsrc cells, treated with 2 µM of the Src inhibitor SU6656 (23), were first incubated with 20 ng/ml PE at 19 °C for 1 h. This allows for efficient binding of PE and its accumulation in the endosomal compartment. The cells were then shifted to 37 °C for various periods of time before a 10-min pulse of cysteine S35, which was immediately followed by cell lysis. Proteins in cell lysates were precipitated, and radioactivity was counted by scintillation. The reference level was measured with control cells treated identically except for the addition of PE. As a control, we checked that the effect on protein synthesis is dependent on an intact Golgi apparatus. This was carried out by treating the cells, in addition to PE, with the drug brefeldin A, which fuses the Golgi apparatus with the ER, therefore inhibiting retrograde transport. Treatment with brefeldin A inhibited the PE-mediated effects on protein synthesis (Fig 4A). To test whether Src expression perturbs the traffic of PE, the cell lines SYF and SYFsrc were compared. Maximal protein synthesis inhibition by PE was obtained after 60 min in SYF cells (Fig 4A). In SYFsrc cells, it took twice as long to reach a similar effect on protein synthesis (Fig 4A), indicating that, in the presence of Src, PE traffic to the ER is slower. Consistently, treatment of SYFsrc cells with the Src inhibitor SU6656, prior to addition of PE, reduced the kinetic of PE effect to a level similar to SYF cells (Fig 4A), indicating that Src-mediated effect on PE transport is direct. To address the possibility that this regulation could occur at the level of endocytosis or endosomal sorting, we followed the intracellular traffic of PE. We labeled PE by covalent attachment of the fluorescent dye Alexa 594® (P3-A594). SYF, Src+, and SYFsrc cells were incubated with PE-A594 at 19 °C for 30 min and then transferred to 37 °C. At 19 °C, PE-A594 accumulated in the endosomes and did not colocalize with the Golgi marker mannosidase II (Fig. 4B). However, after 45 min of incubation at 37 °C, PE began accumulating in the TGN/Golgi compartment in both SYF and SYFsrc, reaching maximum accumulation after 60 min. The amount of PE-A594 that was accumulated in the TGN/Golgi was appreciably lower than the amount detected in the endosomes, suggesting that a significant fraction was degraded in lysosomes. Nevertheless, the kinetics of accumulation of PE-594 in the TGN/Golgi was not significantly different in SYF, Src+, or SYFsrc, indicating that Src does not affect endocytosis or transport of PE from the endosomes to the TGN/Golgi apparatus.

View larger version (28K): [in this window] [in a new window]   FIG. 4. Src expression reduces the rate of Golgi to ER traffic of the Pseudomonas exotoxin. A, SYF, SYF-src, and SYFsrc+ cells treated with SU6656 for 30 min were incubated at 19 °C for 30 min with 20 ng/ml PE before transfer at 37 °C for 15, 30, 60, 90, or 120 min. For each time point, three identical wells of cells were subjected to a 10-min pulse labeling with [35S]methionine. Protein synthesis activity was then measured by scintillation and averaged before normalization to the synthesis in control cells. Control cells were treated identically except for the addition of the toxin. A control including a treatment with brefeldin A at the same time as the toxin shows that the effect of the toxin is dependent on retrograde traffic through the Golgi apparatus. The results shown are representative of three independent experiments. Error bars represent the standard deviation between the three wells. B, to test whether Src expression regulates endocytosis and traffic from endosomes to the Golgi of the exotoxin, PE traffic was followed by microscopy. PE was first covalently labeled with the Alexa 594 fluorophore. The cells were incubated with this fluorescent toxin at 19 °C for 30 min and then fixed (two upper) or shifted for 15', 30', 45', and 60' (two lower rows) at 37 °C before fixation. The cells were then labeled and imaged for mannosidase II (MannII) and the Alexa-594-labeled toxin (PE594).

View larger version (32K): [in this window] [in a new window]   FIG. 5. The deletion of Src does not affect transport of VSV-G protein in the anterograde pathway. Src+ and SYF cells were transfected with GFP-VSVGtsO45. After 24 h, the cells were shifted to 40 °C for 6 h, then cycloheximide was added to the culture medium at 100 µg/ml. Cells were then either fixed (time 0) or shifted to 32 °C for various times up to 90 min. Representative example of cells at 0 min (A, B, C, and D), 20 min (E, F, G, and H), and 90 min (I, J, K, and L) are shown. The cells were labeled with the rabbit polyclonal anti-GRASP 65 to visualize the Golgi (A, E, and I) and imaged for GFP (B, F, and J). The images of the Golgi were used as filters to select, from the GFP image with fluorescence above background, the pixels that were not present in the Golgi region (C, G, and K) and the pixels that were present in the Golgi region (D, H, L). The average intensity (A,in panels C, D, G, H, K, and L) and the ratio (R, in panels D, H, and L) of the fluorescence at the Golgi and in the rest of the cell were then measured. 25 cells for each cell line and for each time point (0, 10, 20, 60, and 90 min) were analyzed, and the results (M) were submitted to statistical analysis; although a statistically significant difference was found between different time points, no difference could be detected between cell lines at a same time point.

	DISCUSSION

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS DISCUSSION REFERENCES

Using the viral protein VSV-GtsO45 as a marker of anterograde traffic, we could not detect any effect on its transport along the secretory pathway upon genetic deletion (Fig. 5) or by pharmacological inhibition of Src (data not shown). This suggests that Src does not play a major role in constitutive transport of secretory protein. We also tested the kinetics of anterograde traffic of Golgi enzymes from the ER upon recovery of a brefeldin A treatment and we found no difference in SYF and SYFsrc cells.² As Src-like kinases are not present in yeast, the Src-dependent regulation of retrograde traffic has to be specific for higher eukaryotes. How Src affects the Golgi organization and specific KDELR dynamics are the obvious challenges given the important oncogenic potential of this kinase.

	FOOTNOTES

 
^* This work was supported by a grant from Ligue Contre le Cancer, Comité du Rhone (to F. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^** These authors contributed equally to this work.

To whom correspondence should be addressed. Tel.: 858-534-3752; Fax: 858-534-0555; E-mail: fabard{at}biomail.ucsd.edu.

¹ The abbreviations used are: SYF, Src, Yes, and Fyn triple knock-out; KDEL-R, KDEL receptor; ER, endoplasmic reticulum; PBS, phosphate-buffered saline; PE, Pseudomonas exotoxin; VSV, vesicular stomatitis virus; GFP, green fluorescent protein; TGN, trans-Golgi network; F-Golgi, GFP fluorescence in the Golgi area; F-ER, GFP fluorescence in the ER.

² F. Bard and P. Jurdic, unpublished observations.

	ACKNOWLEDGMENTS

 
We are very grateful to Drs. Klinghoffer and Soriano for providing the cell lines they generated. We thank the members of Dr. Malhotra's lab for their critical reading of the manuscript.

	REFERENCES

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS DISCUSSION REFERENCES

 

1. Martin, G. S. (2001) Nat. Rev. Mol. Cell Biol. 2, 467–475[CrossRef][Medline] [Order article via Infotrieve]
2. Bjorge, J. D., Jakymiw, A., and Fujita, D. J. (2000) Oncogene 19, 5620–5635[CrossRef][Medline] [Order article via Infotrieve]
3. David-Pfeuty, T., and Nouvian-Dooghe, Y. (1990) J. Cell Biol. 111, 3097–3116[Abstract/Free Full Text]
4. Linstedt, A. D., Vetter, M. L., Bishop, J. M., and Kelly, R. B. (1992) J. Cell Biol. 117, 1077–1084[Abstract/Free Full Text]
5. Foster-Barber, A., and Bishop, J. M. (1998) Proc. Natl. Acad. Sci. U. S. A. 95, 4673–4677[Abstract/Free Full Text]
6. Onofri, F., Giovedi, S., Kao, H. T., Valtorta, F., Borbone, L. B., De Camilli, P., Greengard, P., and Benfenati, F. (2000) J. Biol. Chem. 275, 29857–29867[Abstract/Free Full Text]
7. Onofri, F., Giovedi, S., Vaccaro, P., Czernik, A. J., Valtorta, F., De Camilli, P., Greengard, P., and Benfenati, F. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 12168–12173[Abstract/Free Full Text]
8. Barnekow, A., Jahn, R., and Schartl, M. (1990) Oncogene 5, 1019–1024[Medline] [Order article via Infotrieve]
9. Janz, R., and Sudhof, T. C. (1998) J. Biol. Chem. 273, 2851–2857[Abstract/Free Full Text]
10. Bard, F., Patel, U., Levy, J. B., Horne, W. C., Jurdic, P., and Baron, R. (2002) Eur. J. Cell Biol. 81, 26–35[CrossRef][Medline] [Order article via Infotrieve]
11. Chiu, V. K., Bivona, T., Hach, A., Sajous, J. B., Silletti, J., Wiener, H., Johnson, R. L., Cox, A. D., and Philips, M. R. (2002) Nat. Cell Biol. 4, 343–350[Medline] [Order article via Infotrieve]
12. Jakymiw, A., Raharjo, E., Rattner, J. B., Eystathioy, T., Chan, E. K., and Fujita, D. J. (2000) J. Biol. Chem. 275, 4137–4144[Abstract/Free Full Text]
13. Jamora, C., Takizawa, P. A., Zaarour, R. F., Denesvre, C., Faulkner, D. J., and Malhotra, V. (1997) Cell 91, 617–626[CrossRef][Medline] [Order article via Infotrieve]
14. Liljedahl, M., Maeda, Y., Colanzi, A., Ayala, I., Van Lint, J., and Malhotra, V. (2001) Cell 104, 409–420[CrossRef][Medline] [Order article via Infotrieve]
15. Colanzi, A., Deerinck, T. J., Ellisman, M. H., and Malhotra, V. (2000) J. Cell Biol. 149, 331–339[Abstract/Free Full Text]
16. Acharya, U., Mallabiabarrena, A., Acharya, J. K., and Malhotra, V. (1998) Cell 92, 183–192[CrossRef][Medline] [Order article via Infotrieve]
17. Sutterlin, C., Lin, C. Y., Feng, Y., Ferris, D. K., Erikson, R. L., and Malhotra, V. (2001) Proc. Natl. Acad. Sci. U. S. A. 98, 9128–9132[Abstract/Free Full Text]
18. Thomas, S. M., and Brugge, J. S. (1997) Annu. Rev. Cell Dev. Biol. 13, 513–609[CrossRef][Medline] [Order article via Infotrieve]
19. Klinghoffer, R. A., Sachsenmaier, C., Cooper, J. A., and Soriano, P. (1999) EMBO J. 18, 2459–2471[CrossRef][Medline] [Order article via Infotrieve]
20. Lewis, M. J., and Pelham, H. R. (1992) Cell 68, 353–364[CrossRef][Medline] [Order article via Infotrieve]
21. Jackson, M. E., Simpson, J. C., Girod, A., Pepperkok, R., Roberts, L. M., and Lord, J. M. (1999) J. Cell Sci. 112, 467–475[Abstract]
22. Sandvig, K., and Van Deurs, B. (2002) Annu. Rev. Cell Dev. Biol. 18, 1–24[Medline] [Order article via Infotrieve]
23. Blake, R. A., Broome, M. A., Liu, X., Wu, J., Gishizky, M., Sun, L., and Courtneidge, S. A. (2000) Mol. Cell. Biol. 20, 9018–9027[Abstract/Free Full Text]
24. Griffiths, G., Pfeiffer, S., Simons, K., and Matlin, K. (1985) J. Cell Biol. 101, 949–964[Abstract/Free Full Text]

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				A. Shrivastav, S. Varma, Z. Lawman, S. H. Yang, S. A. Ritchie, K. Bonham, S. M. Singh, A. Saxena, and R. K. Sharma Requirement of N-Myristoyltransferase 1 in the Development of Monocytic Lineage J. Immunol., January 15, 2008; 180(2): 1019 - 1028. [Abstract] [Full Text] [PDF]

				V. Gupta and G. Swarup Evidence for a role of transmembrane protein p25 in localization of protein tyrosine phosphatase TC48 to the ER J. Cell Sci., May 1, 2006; 119(9): 1703 - 1714. [Abstract] [Full Text] [PDF]

				E. J. Tisdale and C. R. Artalejo Src-dependent aProtein Kinase C {iota}/{lambda} (aPKC{iota}/{lambda}) Tyrosine Phosphorylation Is Required for aPKC{iota}/{lambda} Association with Rab2 and Glyceraldehyde-3-phosphate Dehydrogenase on Pre-Golgi Intermediates J. Biol. Chem., March 31, 2006; 281(13): 8436 - 8442. [Abstract] [Full Text] [PDF]

				Y. Posen, V. Kalchenko, R. Seger, A. Brandis, A. Scherz, and Y. Salomon Manipulation of redox signaling in mammalian cells enabled by controlled photogeneration of reactive oxygen species J. Cell Sci., May 1, 2005; 118(9): 1957 - 1969. [Abstract] [Full Text] [PDF]

				P. de Graaf, W. T. Zwart, R. A.J. van Dijken, M. Deneka, T. K.F. Schulz, N. Geijsen, P. J. Coffer, B. M. Gadella, A. J. Verkleij, P. van der Sluijs, et al. Phosphatidylinositol 4-Kinase{beta} Is Critical for Functional Association of rab11 with the Golgi Complex Mol. Biol. Cell, April 1, 2004; 15(4): 2038 - 2047. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF) All Versions of this Article: 278/47/46601    most recent M302221200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Bard, F. Articles by Jurdic, P. Search for Related Content PubMed PubMed Citation Articles by Bard, F. Articles by Jurdic, P. Social Bookmarking                      What's this?