Limits...
Sul1 and Sul2 sulfate transceptors signal to protein kinase A upon exit of sulfur starvation.

Kankipati HN, Rubio-Texeira M, Castermans D, Diallinas G, Thevelein JM - J. Biol. Chem. (2015)

Bottom Line: Overall, our data suggest that transceptors can undergo independent conformational changes, each responsible for triggering different downstream processes.The Sul1 and Sul2 transceptors are the first identified plasma membrane sensors for extracellular sulfate.High affinity transporters induced upon starvation for their substrate may generally act as transceptors during exit from starvation.

View Article: PubMed Central - PubMed

Affiliation: From the Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, Kasteelpark Arenberg 31, B-3001 Leuven-Heverlee, Flanders, Belgium, the Department of Molecular Microbiology, VIB, Kasteelpark Arenberg 31, B-3001 Leuven-Heverlee, Flanders, Belgium, and.

Show MeSH

Related in: MedlinePlus

Uncoupling signaling from endocytosis in sulfate transceptors.A, Western blot detection of immunoprecipitated HA-tagged Sul proteins. Shown are Sul1-HA or Sul2-HA in membrane-enriched P13 fractions isolated from cells before and after the addition of 3 mm sulfate or 3 mmd-glucosamine 2-sulfate. Shown are Sul1E427Q-HA and Sul2E443Q-HA before and after the addition of 3 mm sulfate. B and C, localization of HA-tagged Sul proteins as detected by immunofluorescence and confocal microscopy imaging. B, Sul1-HA or Sul2-HA before and after the addition of 3 mmd-glucosamine 2-sulfate. C, Sul1E427Q-HA or Sul2E443Q-HA before and after the addition of 3 mm sulfate. In all immunofluorescence experiments, HA-tagged Sul proteins were detected with anti-HA rat primary antibody and Alexa Fluor 488-conjugated anti-rat secondary antibody. DIC, differential interference contrast.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4400352&req=5

Figure 8: Uncoupling signaling from endocytosis in sulfate transceptors.A, Western blot detection of immunoprecipitated HA-tagged Sul proteins. Shown are Sul1-HA or Sul2-HA in membrane-enriched P13 fractions isolated from cells before and after the addition of 3 mm sulfate or 3 mmd-glucosamine 2-sulfate. Shown are Sul1E427Q-HA and Sul2E443Q-HA before and after the addition of 3 mm sulfate. B and C, localization of HA-tagged Sul proteins as detected by immunofluorescence and confocal microscopy imaging. B, Sul1-HA or Sul2-HA before and after the addition of 3 mmd-glucosamine 2-sulfate. C, Sul1E427Q-HA or Sul2E443Q-HA before and after the addition of 3 mm sulfate. In all immunofluorescence experiments, HA-tagged Sul proteins were detected with anti-HA rat primary antibody and Alexa Fluor 488-conjugated anti-rat secondary antibody. DIC, differential interference contrast.

Mentions: We next investigated whether signaling and transport could be uncoupled from endocytosis using the non-transported signaling agonist, d-glucosamine 2-sulfate. The addition of d-glucosamine 2-sulfate did not cause any disappearance of Sul1-HA or Sul2-HA in immunoprecipitates of membrane enriched P13 fractions (Fig. 8A). This was confirmed with immunofluorescence microscopy, which showed no detectable change in the intracellular localization of Sul1-HA and Sul2-HA up to 240 min after the addition of d-glucosamine 2-sulfate (Fig. 8B). These results show that signaling is either not a trigger or at least not sufficient to trigger endocytosis.


Sul1 and Sul2 sulfate transceptors signal to protein kinase A upon exit of sulfur starvation.

Kankipati HN, Rubio-Texeira M, Castermans D, Diallinas G, Thevelein JM - J. Biol. Chem. (2015)

Uncoupling signaling from endocytosis in sulfate transceptors.A, Western blot detection of immunoprecipitated HA-tagged Sul proteins. Shown are Sul1-HA or Sul2-HA in membrane-enriched P13 fractions isolated from cells before and after the addition of 3 mm sulfate or 3 mmd-glucosamine 2-sulfate. Shown are Sul1E427Q-HA and Sul2E443Q-HA before and after the addition of 3 mm sulfate. B and C, localization of HA-tagged Sul proteins as detected by immunofluorescence and confocal microscopy imaging. B, Sul1-HA or Sul2-HA before and after the addition of 3 mmd-glucosamine 2-sulfate. C, Sul1E427Q-HA or Sul2E443Q-HA before and after the addition of 3 mm sulfate. In all immunofluorescence experiments, HA-tagged Sul proteins were detected with anti-HA rat primary antibody and Alexa Fluor 488-conjugated anti-rat secondary antibody. DIC, differential interference contrast.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4400352&req=5

Figure 8: Uncoupling signaling from endocytosis in sulfate transceptors.A, Western blot detection of immunoprecipitated HA-tagged Sul proteins. Shown are Sul1-HA or Sul2-HA in membrane-enriched P13 fractions isolated from cells before and after the addition of 3 mm sulfate or 3 mmd-glucosamine 2-sulfate. Shown are Sul1E427Q-HA and Sul2E443Q-HA before and after the addition of 3 mm sulfate. B and C, localization of HA-tagged Sul proteins as detected by immunofluorescence and confocal microscopy imaging. B, Sul1-HA or Sul2-HA before and after the addition of 3 mmd-glucosamine 2-sulfate. C, Sul1E427Q-HA or Sul2E443Q-HA before and after the addition of 3 mm sulfate. In all immunofluorescence experiments, HA-tagged Sul proteins were detected with anti-HA rat primary antibody and Alexa Fluor 488-conjugated anti-rat secondary antibody. DIC, differential interference contrast.
Mentions: We next investigated whether signaling and transport could be uncoupled from endocytosis using the non-transported signaling agonist, d-glucosamine 2-sulfate. The addition of d-glucosamine 2-sulfate did not cause any disappearance of Sul1-HA or Sul2-HA in immunoprecipitates of membrane enriched P13 fractions (Fig. 8A). This was confirmed with immunofluorescence microscopy, which showed no detectable change in the intracellular localization of Sul1-HA and Sul2-HA up to 240 min after the addition of d-glucosamine 2-sulfate (Fig. 8B). These results show that signaling is either not a trigger or at least not sufficient to trigger endocytosis.

Bottom Line: Overall, our data suggest that transceptors can undergo independent conformational changes, each responsible for triggering different downstream processes.The Sul1 and Sul2 transceptors are the first identified plasma membrane sensors for extracellular sulfate.High affinity transporters induced upon starvation for their substrate may generally act as transceptors during exit from starvation.

View Article: PubMed Central - PubMed

Affiliation: From the Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, Kasteelpark Arenberg 31, B-3001 Leuven-Heverlee, Flanders, Belgium, the Department of Molecular Microbiology, VIB, Kasteelpark Arenberg 31, B-3001 Leuven-Heverlee, Flanders, Belgium, and.

Show MeSH
Related in: MedlinePlus