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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:24818*  Cite 
Description

Intracellular localization of GFP-Vps74 K178A, R181A mutant in BY4742 sac1∆ vps74∆ mutant. SAC1 encodes an integral membrane phosphoinositide phosphatase and in the sac1∆ vps74∆ mutant, GFP-Vps74 localizes, in addition to the Golgi apparatus, to nuclear ER and cortical ER and/or PM (CIL# 24816). This localization of GFP-Vps74 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 K178A R181A/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.

Biological Sources
NCBI Organism Classification
Saccharomyces cerevisiae S288c
Cell Line
BY4742 sac1∆ vps74∆
Cellular Component
Golgi apparatus
cytosol
extrinsic to membrane
nucleus
integral to endoplasmic reticulum membrane
integral to Golgi membrane
Attribution
Names
Christopher S. Wood
Karl R. Schmitz
Nicholas J. Bessman
Thanuja Gangi Setty
Kathryn M. Ferguson
Christopher G. Burd
Published
J Cell Biol. 187: 967-975. 2009
Pubmed
20026658
Link
http://jcb-dataviewer.rupress.org/jcb/browse/...
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL24818
Archival Resource Key (ARK)
ark:/b7295/w9cil24818
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
fluorescence microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
EGFP
Processing History
constrained iterative deconvolution
Data Qualifiers
processed data
suitable for spatial measurements
Sample Preparation
Methods
living tissue
Relation To Intact Cell
whole mounted tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 302px 0.0663µm
Y 302px 0.0663µm
Z 14px 0.4µm