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Description
Intracellular localization of GFP-GOLPH3 in BY4742 vps74∆ sac1∆. SAC1 encodes an integral membrane phosphoinositide phosphatase and in the sac1∆ vps74∆ mutant, GFP-GOLPH3 localizes, in addition to the Golgi apparatus, to nuclear ER and cortical ER and/or PM. Control image for images showing ectopic expression of GFP-GOLPH3 with mutations in the sulfate-binding pocket (CIL#s 24820, 24821). This study demonstrates that the sulfate-binding pocket of Vps74 and GOLPH3 mediates PtdIns4P-binding and is essential for function. 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 Fig4A GFP-GOLPH3 wild-type/sac1∆ panel in J Cell Biol. 187: 967-975. 2009. Images in Fig 4A include CIL#s 13449, 13451, 13453, 13455, 13456, 13457, 24816, 24817, 24818, 24819, 24820, 24821. 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:24819, Saccharomyces cerevisiae S288c. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J08K77XF
Type
image
Identifier
ark:/20775/bb3658569s
Language
No linguistic content
Subject
Protein transport Phosphatidylinositol-4-phosphate binding Retrograde vesicle-mediated transport, Golgi to ER BY4742 vps74∆ sac1∆ Enzyme binding Phosphatidylinositol dephosphorylation Protein localization in Golgi apparatus Phosphatidylinositol-4-phosphate phosphatase activity Nucleus Integral to endoplasmic reticulum membrane Golgi apparatus Cytosol Integral to Golgi membrane Extrinsic to membrane Saccharomyces cerevisiae S288c Cell Image Library Group ID: 6596
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