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Description
Intracellular localization of GFP-GOLPH3 R171A, R174A mutant in BY4742 sac1∆ vps74∆ mutant. 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 (CIL# 24819). This localization of GFP-GOLPH3 is lost when two amino acids are mutated in the sulfate-binding pocket. 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 R171A R174A/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:24821, Saccharomyces cerevisiae S288c. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0D7995Z
Type
image
Identifier
ark:/20775/bb5365068r
Language
No linguistic content
Subject
BY4742 sac1∆ vps74∆ Phosphatidylinositol-4-phosphate phosphatase activity Phosphatidylinositol dephosphorylation Protein transport Phosphatidylinositol-4-phosphate binding Retrograde vesicle-mediated transport, Golgi to ER Protein localization in Golgi apparatus Enzyme binding Golgi apparatus Nucleus Integral to Golgi membrane Cytosol Integral to endoplasmic reticulum membrane Extrinsic to membrane Saccharomyces cerevisiae S288c Cell Image Library Group ID: 6596
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