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Transport into and out of the Golgi complex studied by transfecting cells with cDNAs encoding horseradish peroxidase
- Source :
- The Journal of Cell Biology
- Publication Year :
- 1994
- Publisher :
- Rockefeller University Press, 1994.
-
Abstract
- We have developed a novel technique with which to investigate the morphological basis of exocytotic traffic. We have used expression of HRP from cDNA in a variety of cells in combination with peroxidase cytochemistry to outline traffic into and out of the Golgi apparatus at the electron microscopic level with very high sensitivity. A secretory form of the peroxidase (ssHRP) is active from the beginning of the secretory pathway and the activity is efficiently cleared from cells. Investigation of the morphological elements involved in the itinerary of soluble ER proteins using ssHRP tagged with the ER retention motif (ssHRPKDEL) shows that it progresses through the Golgi stack no further than the cis-most element. Traffic between the RER and the Golgi stack as outlined by ssHRPKDEL occurs via vesicular carriers as well as by tubular elements. ssHRP has also been used to investigate the trans side of the Golgi complex, where incubation at reduced temperatures outlines the trans-Golgi network with HRP reaction product. Tracing the endosomal compartment with transferrin receptor in double-labeling experiments with ssHRP fails to show any overlap between these two compartments.
- Subjects :
- Carcinoma, Hepatocellular
DNA, Complementary
Endosome
Molecular Sequence Data
Genes, myc
Golgi Apparatus
Endosomes
Endoplasmic Reticulum
Kidney
Transfection
Polymerase Chain Reaction
Horseradish peroxidase
Cell Line
symbols.namesake
chemistry.chemical_compound
Chlorocebus aethiops
Receptors, Transferrin
Animals
Humans
Horseradish Peroxidase
Secretory pathway
DNA Primers
Base Sequence
biology
Endoplasmic reticulum
Liver Neoplasms
ER retention
Articles
Cell Biology
Golgi apparatus
Brefeldin A
Endocytosis
Cell biology
Kinetics
Secretory protein
chemistry
symbols
biology.protein
Subjects
Details
- ISSN :
- 15408140 and 00219525
- Volume :
- 127
- Database :
- OpenAIRE
- Journal :
- Journal of Cell Biology
- Accession number :
- edsair.doi.dedup.....61b0ba8ca914b4bcf5573fd9abede1ad