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A genome-wide screen for genes affecting eisosomes reveals Nce102 function in sphingolipid signaling.

Fröhlich F, Moreira K, Aguilar PS, Hubner NC, Mann M, Walter P, Walther TC - J. Cell Biol. (2009)

Bottom Line: The relative abundance of Nce102 in these domains compared with the rest of the plasma membrane is dynamically regulated by sphingolipids.Furthermore, Nce102 inhibits Pkh kinase signaling and is required for plasma membrane organization.Therefore, Nce102 might act as a sensor of sphingolipids that regulates plasma membrane function.

View Article: PubMed Central - PubMed

Affiliation: Organelle Architecture and Dynamics, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany.

ABSTRACT
The protein and lipid composition of eukaryotic plasma membranes is highly dynamic and regulated according to need. The sphingolipid-responsive Pkh kinases are candidates for mediating parts of this regulation, as they affect a diverse set of plasma membrane functions, such as cortical actin patch organization, efficient endocytosis, and eisosome assembly. Eisosomes are large protein complexes underlying the plasma membrane and help to sort a group of membrane proteins into distinct domains. In this study, we identify Nce102 in a genome-wide screen for genes involved in eisosome organization and Pkh kinase signaling. Nce102 accumulates in membrane domains at eisosomes where Pkh kinases also localize. The relative abundance of Nce102 in these domains compared with the rest of the plasma membrane is dynamically regulated by sphingolipids. Furthermore, Nce102 inhibits Pkh kinase signaling and is required for plasma membrane organization. Therefore, Nce102 might act as a sensor of sphingolipids that regulates plasma membrane function.

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Nce102 localizes to both MCC and non-MCC domains in the plasma membrane. (a) Images from cells expressing Nce102-GFP (left; green in overlay) and Sur7-mars (middle; red in the overlay) are shown. (b) Intensity profiles of Nce102-GFP and Sur7-mars along the plasma membrane. (c) Images from cells expressing Nce102-GFP (left; green in overlay) and Lsp1-mars (middle; red in the overlay) are shown. Boxes indicate the area magnified in the bottom panels. (d) Intensity profiles of Nce102-GFP and Lsp1-mars along the plasma membrane. (e) Pil1 is required for normal Nce102 distribution. Wild-type (left) and Δpil1 (right) cells expressing Nce102-GFP are shown. Arrows highlight eisosome remnants. Bars, 5 µm.
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fig3: Nce102 localizes to both MCC and non-MCC domains in the plasma membrane. (a) Images from cells expressing Nce102-GFP (left; green in overlay) and Sur7-mars (middle; red in the overlay) are shown. (b) Intensity profiles of Nce102-GFP and Sur7-mars along the plasma membrane. (c) Images from cells expressing Nce102-GFP (left; green in overlay) and Lsp1-mars (middle; red in the overlay) are shown. Boxes indicate the area magnified in the bottom panels. (d) Intensity profiles of Nce102-GFP and Lsp1-mars along the plasma membrane. (e) Pil1 is required for normal Nce102 distribution. Wild-type (left) and Δpil1 (right) cells expressing Nce102-GFP are shown. Arrows highlight eisosome remnants. Bars, 5 µm.

Mentions: To answer how Nce102 functions, we first investigated its subcellular localization. In agreement with a recent study that identified Nce102 as an MCC component, we found Nce102-GFP localizing in the plasma membrane and accumulating in foci reminiscent of MCCs (Fig. 3 a; Grossmann et al., 2008). This notion was further confirmed by colocalization of Nce102 with plasma membrane markers but not, for example, with cortical ER markers (unpublished data).


A genome-wide screen for genes affecting eisosomes reveals Nce102 function in sphingolipid signaling.

Fröhlich F, Moreira K, Aguilar PS, Hubner NC, Mann M, Walter P, Walther TC - J. Cell Biol. (2009)

Nce102 localizes to both MCC and non-MCC domains in the plasma membrane. (a) Images from cells expressing Nce102-GFP (left; green in overlay) and Sur7-mars (middle; red in the overlay) are shown. (b) Intensity profiles of Nce102-GFP and Sur7-mars along the plasma membrane. (c) Images from cells expressing Nce102-GFP (left; green in overlay) and Lsp1-mars (middle; red in the overlay) are shown. Boxes indicate the area magnified in the bottom panels. (d) Intensity profiles of Nce102-GFP and Lsp1-mars along the plasma membrane. (e) Pil1 is required for normal Nce102 distribution. Wild-type (left) and Δpil1 (right) cells expressing Nce102-GFP are shown. Arrows highlight eisosome remnants. Bars, 5 µm.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2712959&req=5

fig3: Nce102 localizes to both MCC and non-MCC domains in the plasma membrane. (a) Images from cells expressing Nce102-GFP (left; green in overlay) and Sur7-mars (middle; red in the overlay) are shown. (b) Intensity profiles of Nce102-GFP and Sur7-mars along the plasma membrane. (c) Images from cells expressing Nce102-GFP (left; green in overlay) and Lsp1-mars (middle; red in the overlay) are shown. Boxes indicate the area magnified in the bottom panels. (d) Intensity profiles of Nce102-GFP and Lsp1-mars along the plasma membrane. (e) Pil1 is required for normal Nce102 distribution. Wild-type (left) and Δpil1 (right) cells expressing Nce102-GFP are shown. Arrows highlight eisosome remnants. Bars, 5 µm.
Mentions: To answer how Nce102 functions, we first investigated its subcellular localization. In agreement with a recent study that identified Nce102 as an MCC component, we found Nce102-GFP localizing in the plasma membrane and accumulating in foci reminiscent of MCCs (Fig. 3 a; Grossmann et al., 2008). This notion was further confirmed by colocalization of Nce102 with plasma membrane markers but not, for example, with cortical ER markers (unpublished data).

Bottom Line: The relative abundance of Nce102 in these domains compared with the rest of the plasma membrane is dynamically regulated by sphingolipids.Furthermore, Nce102 inhibits Pkh kinase signaling and is required for plasma membrane organization.Therefore, Nce102 might act as a sensor of sphingolipids that regulates plasma membrane function.

View Article: PubMed Central - PubMed

Affiliation: Organelle Architecture and Dynamics, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany.

ABSTRACT
The protein and lipid composition of eukaryotic plasma membranes is highly dynamic and regulated according to need. The sphingolipid-responsive Pkh kinases are candidates for mediating parts of this regulation, as they affect a diverse set of plasma membrane functions, such as cortical actin patch organization, efficient endocytosis, and eisosome assembly. Eisosomes are large protein complexes underlying the plasma membrane and help to sort a group of membrane proteins into distinct domains. In this study, we identify Nce102 in a genome-wide screen for genes involved in eisosome organization and Pkh kinase signaling. Nce102 accumulates in membrane domains at eisosomes where Pkh kinases also localize. The relative abundance of Nce102 in these domains compared with the rest of the plasma membrane is dynamically regulated by sphingolipids. Furthermore, Nce102 inhibits Pkh kinase signaling and is required for plasma membrane organization. Therefore, Nce102 might act as a sensor of sphingolipids that regulates plasma membrane function.

Show MeSH