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Description
Intracellular localization of GFP-GOLPH3 in BY4742 sac1∆ vps74∆. SAC1 encodes an integral membrane phosphoinositide phosphatase that is localized to the ER and Golgi. In the sac1∆ vps74∆ mutant, GFP-GOLPH3 localizes, in addition to the Golgi apparatus, to nuclear ER and cortical ER and/or PM. Thus, correct Golgi localization of Vps74 (see CIL# 24812 for GFP-Vps74 in sac1∆ vps74∆ mutant) and GOLPH3 requires synthesis of PtdIns4P in the Golgi by Pik1-Frq1 and maintenance of low PtdIns4P levels in non-Golgi membranes by the Sac1 lipid phosphatase. Cells grown in liquid medium were mounted in growth medium and 3D image stacks were collected at 0.4-µm z increments on a DeltaVision workstation (Applied Precision) based on an inverted microscope (IX-70; Olympus) using a 100× NA 1.4 oil immersion lens. Images were captured at 23C with a 12-bit CCD camera (CoolSnap HQ; Photometrics) and deconvolved using the iterative-constrained algorithm (Agard, 1984) and the measured point spread function. One image from the approximate center of z stack is shown in Fig1B GOLPH3/sac1∆ panel in J Cell Biol. 187: 967-975. 2009. Images in Fig 1B include CIL#s 24815, 24812, 24813. Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp) Wood, Christopher S.; Schmitz, Karl R.; Bessman, Nicholas J.; Setty, Thanuja Gangi; Ferguson, Kathryn M.; Burd, Christopher G. (2021). CIL:24815, Saccharomyces cerevisiae S288c. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0D7999R
Type
image
Identifier
ark:/20775/bb71056971
Language
No linguistic content
Subject
BY4742 sac1∆ vps74∆ Enzyme binding Retrograde vesicle-mediated transport, Golgi to ER Phosphatidylinositol-4-phosphate binding Protein localization in Golgi apparatus Phosphatidylinositol-4-phosphate phosphatase activity Phosphatidylinositol dephosphorylation Protein transport Golgi apparatus Integral to endoplasmic reticulum membrane Extrinsic to membrane Endoplasmic reticulum Integral to Golgi membrane Nucleus Cytosol Saccharomyces cerevisiae S288c Cell Image Library Group ID: 6596
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