Retention of secretory proteins in an intermediate compartment and disappearance of the Golgi complex in an END4 mutant of Chinese hamster ovary cells.
Mutant V.24.1, a member of the End4 complementation group of temperature-sensitive CHO cells, is defective in secretion at the restrictive temperature (Wang, R.-H., P. A. Colbaugh, C.-Y. Kao, E. A. Rutledge, and R. K. Draper. 1990. J. Biol. Chem. 265:20179-20187; Presley, J. F., R. K. Draper, and D. T. Brown. 1991. J. Virol. 65:1332-1339). We have further investigated the secretory lesion and report three main findings. First, the block in secretion is not due to aberrant folding or oligomerization of secretory proteins in the endoplasmic reticulum because the hemagglutinin of influenza virus folded and oligomerized at the same rate in mutant and parental cells at the restrictive temperature. Second, secretory proteins accumulated in a compartment intermediate between the ER and the Golgi. Several lines of evidence support this conclusion, the most direct being the colocalization by immunofluorescence microscopy of influenza virus hemagglutinin with a 58-kD protein that is known to reside in an intermediate compartment. Third, at the resolution of fluorescence microscopy, the Golgi complex in the mutant cells vanished at the restrictive temperature.
Kao
C Y
CY
Molecular and Cell Biology Program, University of Texas, Dallas, Richardson 75083-0688.
Draper
R K
RK
D000818:Animals; D001692:Biological Transport; D020126:Brefeldin A; D016466:CHO Cells; D002451:Cell Compartmentation; D006224:Cricetinae; D003517:Cyclopentanes / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism*; D006389:Hemagglutinins, Viral / Q000378:metabolism; D006596:Hexosaminidases / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000378:metabolism; D008870:Microtubules / Q000502:physiology; D008985:Monensin / Q000494:pharmacology; D009154:Mutation; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism*; D013696:Temperature; D013997:Time Factors
D003517:Cyclopentanes; D006023:Glycoproteins; D006389:Hemagglutinins, Viral; D046911:Macromolecular Substances; D008565:Membrane Proteins; D011506:Proteins; C034241:ribophorin; D020126:Brefeldin A; D008985:Monensin; D006596:Hexosaminidases